issn 1443-0193 australian biochemist - asbmb · each letter codes for another letter – to get you...

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST

ISSN 1443-0193

THIS ISSUE INCLUDES

SHOWCASE ON RESEARCHLactation

Australian BiochemistThe Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.

Volume 48 APRIL 2017 No.1

Showcase on Research Appetite Control

Hormones in Human Milk Cells in Human Milk –

What Do They Tell Us Human Milk Bioactivity Pesticides in Human Milk

Other Features Great Expectations SDS (Students) Page Competition Travel Report FAOBMB Intellectual Property

Profiles of ASBMB Medallists and AwardeesINSIDE

Science Talent Search Honours for Members Special Interest Groups Our Sustaining Members Forthcoming Meetings Directory

Stem cell

Common progenitor

Myoepithelial progenitor

Myoepithelial Alveolar Ductal

Luminal progenitor

Figure 1

Vol 48 No 1 April 2017AUSTRALIAN BIOCHEMIST

We have another competition for the readers of the Australian Biochemist. All correct entries received by the Editor (email [email protected]) before 14 May 2017 will enter the draw to receive a gift voucher. With thanks to Rebecca Lew.

‘Chemical Cryptogram’ Competition

Solve the coded words below to reveal a special list, described in the underlined title.Each letter codes for another letter – to get you started,the coded letters RPAUMS = BLONDE. The dates are hints.

DAYSU RBAFQSYBJEJ DQA DAU UARSP KCBLSJ

1. VSCET FACB (1947)

2. MACAEQT QAMVGBU (1964)

3. VSCECXMS SPBAU (1988)

4. SPBLIRSEQ RPIFGRXCU (2009)

5. IMI TAUIEQ (2009)

Showcase on ResearchGreat ExpectationsTechnical FeaturesSDS (Students) Page

Coverage of all issues from 2000 to the present


Australian Biochemist - Editor Chu Kong Liew, Editorial Officer Liana Friedman© 2017 Australian Society for Biochemistry and Molecular Biology Inc. All rights reserved.

From the Editor’ s DeskAs we head towards Autumn, it isn’t just the leaves and the weather that are

changing; change is also coming to the Australian Biochemist, in the form of a new editorial team. Beginning with the next issue, Suresh Mathivanan and Tatiana Soares da Costa will be taking over the reins from me. As you well know, the Australian Biochemist has already undergone some changes, most notably the transition to an electronic only format. Under the guidance of Suresh and Tatiana, I have no doubt that the magazine will improve by leaps and bounds, as they take the magazine further forward into the digital era. Suresh’s expertise with technology (evident from his leadership in revamping the ASBMB website) combined with Tatiana’s fresh perspective (having written our Short Discussions for Students Page for the last few years) will ensure that the magazine remains a relevant and an effective tool to meet the communication needs of our membership.

As part of these changes, this issue will feature our very last Showcase on Research, a feature that has been running in the magazine for at least 19 years. Thankfully, we will be finishing on a high note, with a fascinating feature on human breastmilk. Put together by Donna Geddes and her team of authors, the Showcase will explore several topics surrounding this powerhouse solution of nutrients, cells and bioactive molecules, which has effects not only on the developing infant but also on its life further down the track. One feature that isn’t going anywhere (at least for now) is Great Expectations, with this issue’s feature written by Rebecca Lew. As a former Editor of the Australian Biochemist, Rebecca is certainly used to organising and editing Great Expectations but in this issue, she proves herself equally adept at writing an interesting piece too! In her feature, Becky takes us on her journey from biology-loving child in the US to senior lecturer at Monash University and now a senior medical writer.

Following on from her article on the considerations behind filing a patent, Sarah Hennebry describes the sequence of events that take place once a patent been filed. Tatiana Soares da Costa shares some valuable advice on writing and publishing a manuscript. Rounding off this issue are the profiles of the ASBMB award winners, travel award and Special Interest Group reports, and Australia Day Honours for ASBMB members.

As I step down from my role, I’d like to acknowledge all the wonderful help that I’ve received from the Editorial Committee during my time as editor. First and foremost, I’d like to thank Liana Friedman, our Editorial Officer, for her skill, patience and dedication to putting out the highest quality publication with every issue. It is difficult to overstate her contribution to the success of the magazine but I know that with Liana committed to staying on, the Australian Biochemist is in great hands! Thanks go to Rebecca Lew, who introduced me to the job and has provided valuable guidance over the years. I’d also like to thank the other members of the Editorial Committee, both past and present for kindly giving up their time to the magazine. Many thanks also to Sally and Chris Jay, particularly to Sally for her tireless efforts in serving as the nexus between the magazine and our Sustaining Members. Thanks also to the ASBMB Executive Council for their continual help and advice. Last but not least, a big thank you to all our contributors, who generously provide the wonderful content that fills these pages.

Chu Kong Liew

Chu Kong LiewEditor Australian Biochemist

Would you like to help out with the Australian Biochemist?

Then please join our Editorial Committee! If you have an interest in being part of the magazine, we warmly welcome

your involvement. To volunteer or to find out more about what is involved, please contact the Editor at [email protected].


SHOWCASE ON RESEARCH

Lactation in the LimelightED ITOR IAL

Recently, there has been widespread recognition among the scientific community of the programming of long-term health outcomes during pregnancy and the early years of life. Indeed, it is believed that the first 1000 days of life is the most sensitive window of human development. Epidemiologic and interventional animal models provide compelling evidence that nutrition profoundly affects organ development and metabolism, with consequences extending into adult life.

In a landmark series recently published in The Lancet, an extensive meta analysis confirms the significant advantages of breastfeeding, which include decreased childhood infections and malocclusion, likely reductions in overweight and diabetes later in life along, as well as increased intelligence (1). However, the mechanisms by which breastfeeding confer advantages remain to be elucidated.

Further, whilst the benefits to the infant are often cited to encourage breastfeeding, the lactating woman also reaps substantial health benefits, such as decreased risk of breast and ovarian cancer and type 2 diabetes (1). It has been predicted that if breastfeeding were to be scaled up to near universal levels, 823,000 deaths of children under five years of age would be prevented, along with 20,000 deaths from breast cancer per year (1). Finally, human milk and breastfeeding are moving into the spotlight as researchers try to uncover aspects of breastfeeding that confer benefits; these aspects include milk components and the very act of feeding and whether these are affected by maternal characteristics. Such knowledge may potentially provide windows of opportunity to improve nutrition and development of our infants.

This Showcase on Research will explore human milk to a much greater extent than previously documented, due to recent research using cutting edge technology, allowing greater insight into the intriguing intricacies of this complex fluid.

Many of the benefits of breastfeeding are attributed in part to the reduced growth rate of breastfed infants compared to formula-fed infants. Given that breastfed

infants self-regulate their milk intake and that human milk contains bioactive appetite control factors, it is compelling to deduce that both the composition of milk and the volume consumed may be related to infant growth and body composition. Gridneva and Geddes explore this scenario with a complex analysis of human milk composition, carried out with respect to gastric emptying (a key appetite regulator), infant growth and body composition over the first year of life.

Often overlooked is the fact that human milk is a ‘live potion’ that contains amongst its bioactive components, live cells. Most recently, immune cells in milk have been shown to increase in response to both infant and maternal infection, highlighting the intimate relationship between the mother and infant. Further, a full cellular hierarchy has also been identified, with human milk stem cells being shown to be pluripotent and able to develop into numerous cells types when placed in stimulating environments. Twigger and Hartmann discuss the potential of these cells in helping to elucidate the development of the human mammary gland during lactation as well as the possibile discovery of biomarkers for dysfunction and disease.

Sharp et al. discuss the fascinating evolution of the mammary gland and development of the young with respect to the bioactive components of milk. Further, they explore gene expression in the milk of women with mastitis, which is one of the major causes of early weaning in breastfeeding women.

Unfortunately, not all components of milk are considered beneficial for the infant, in particular contaminants present in our environment. Gay et al. articulate the development of a highly sensitive assay to detect persistent organic pesticides in human milk. Further, they calculate rather than estimate the actual dose ingested by the infant over the first year of life and determine if this is related to infant growth.

Enjoy this update of one of the most crucial physiological processes for the survival of the human race!1. Victora, C.G., et al. (2016) Lancet 387, 475-490

ED ITOR IAL

LactationGuest Editor: Donna Geddes5 Appetite Control Hormones in Human Milk – What is Their Role Zoya Gridneva and Donna Geddes 9 Cells in Human Milk – What Do They Tell Us Alecia-Jane Twigger and Peter Hartmann13 Human Milk Bioactivity: Lessons from the Evolution of Lactation

Julie Sharp, Ashalyn Watt, Christophe Lefevre and Kevin Nicholas19 Pesticides in Human Milk: Should We Be Concerned? Melvin Gay, Robert Trengove and Donna Geddes

Donna GeddesSchool of Chemistry and Biochemistry, University of Western Australia, Perth WA 6009 [email protected]

Cover IllustrationMammary stem and differentiated mammary epithelial cells isolated from human milk.Image courtesy of Alecia-Jane Twigger and Peter Hartmann, School of Chemistry and Biochemistry, University of Western Australia.



Appetite Control Hormones in Human Milk –What is Their Role

Zoya Gridneva* and Donna Geddes School of Chemistry and Biochemistry, University of Western Australia, Perth, WA 6009

*Corresponding authors: [email protected]

Demand the BestAll parents seek optimal nutrition for their children and

whilst we believe most know that human milk (HM) meets all the needs of the infant (1), not everyone understands the form of delivery, that is breastfeeding on demand, is also desirable. Breastfeeding mothers meet on average seven deadlines per day in the first months of baby’s life (2) and these deadlines may happen anywhere and anytime. Modern society is still not keen on breastfeeding in public, and any mother daring to address the demand may be frowned upon, yet it is not the actions of a spoiled child we are observing, but the amazing biochemistry behind the appetite control in action.

Human Milk Appetite HormonessBreastfed infants eat less but more frequently than

formula-fed infants and are leaner as a result of lower energy intake (3). They also display a variety of feeding patterns and frequency (2) which can, in part, be explained by HM composition.

HM is a complex mixture consisting not only of nutrients but also bioactive molecules including hormones, growth factors, neuropeptides, immunomodulating and anti-inflammatory agents (4). An increasing number of appetite hormones have been recently identified in HM and a number of studies aimed to elucidate their effect on the infant. These HM appetite hormones are mainly derived from the maternal circulation, with a small contribution by the breast (5,6). As mediators between the adipose tissue, gastrointestinal tract and infant brain, the potential functions of appetite hormones include regulation of satiety, development of appetite control pathways, and modulation of infant growth and development (7) . This review discusses the effect of the appetite hormones on infant gastric emptying (GE) and body composition (BC), as both have important roles in regulating appetite, food intake and energy balance.

Leptin Leptin is a polypeptide hormone synthesised by the

white adipose tissue and is the most widely studied of all appetite hormones. It is involved in the regulation of adipose tissue, food intake and body weight. Leptin causes weight loss by suppressing appetite via signalling satiety and increasing metabolic rate. Leptin in HM is derived mainly from maternal serum, following secretion from white adipocytes and gastric chief cells into the bloodstream, with a small contribution from the mammary gland epithelium (5,8).

In infants, HM is believed to be a major source of leptin early in life, due to immature endogenous leptin-

synthesising mechanisms (9). Indeed, breastfed infants exhibit higher serum leptin levels compared to formula-fed infants, which is likely due to leptin being rendered inactive during the processing of bovine milk for formula (10). Leptin in HM has been hypothesised to be involved both in the short-term control of appetite and in developmental programming of appetite and energy signalling pathways, promoting efficient energy control and storage throughout life (8,11). In support of this, leptin administered during the first 14 days of life acts as a neurotrophic agent, promoting neural growth from the arcuate nucleus of the hypothalamus to multiple appetite control centres located in the central nervous system (12). However the influence of leptin on GE, a key regulator of appetite, has not been well studied. Animal models show that central administration of leptin delays GE (13) and reduces food intake (14). In contrast, we have measured infants’ stomachs (Fig. 1) sequentially to assess the effect of an array of HM components on GE for a single breastfeed and we found that HM leptin does not appear to impact GE in term infants (15,16), nor has it been linked to either the frequency of feeding or time between the feeds (15-17). This suggests the action of HM leptin on short-term appetite control may be mediated by upregulation of circulating melanocortins, potent anorexigenic agents that promote satiety (18). Thus actions other than regulation of GE or downstream effects of HM leptin not yet measured are likely responsible for the beneficial effects of HM in protecting against obesity later in life.

With respect to regulation of BC, higher HM leptin levels are associated with lower infant weight gain in the first six months, lower infant BMI at two years of age (18),

Fig. 1. Ultrasound technique for measuring infant’s stomach. The longitudinal and transverse planes for measurement of stomach volume are indicated.


lower infant weight and adiposity (18,19) and greater lean body mass (total body water) in breastfed infants (20,21), suggesting a pivotal role for leptin in regulating infant growth and BC. Research in our laboratory confirms these results and also shows that both higher HM leptin concentration and 24-hour intake of leptin are associated with lower infant weight, lean body mass and fat mass (measured with bioimpedance spectroscopy, Fig. 2) during first 12 months of lactation (Gridneva and Geddes, unpublished data). Given that many of the benefits of breastfeeding are associated with the growth rate of the infant, these findings suggest that HM is indeed bioactive. Further studies should incorporate exploration of the regulation of leptin levels in HM, which have been shown to be influenced by maternal adiposity, but also display a circadian rhythm (17).

AdiponectinAdiponectin is the appetite hormone present in the

highest concentrations in HM and its concentration is more than 40 times higher than that of ghrelin and leptin (22,23). Adiponectin is secreted by adipose tissue and also synthesised by the breast (5). Adiponectin circulates as oligomers of different sizes, from low-molecular-weight trimers to high-molecular-weight octodecamers, the latter being the most common in HM (24), in contrast to the middle-molecular-weight form of bovine adiponectin (25). It is present in a biologically active form and is resistant to digestion (26). Interestingly, HM adiponectin levels are only moderately related to both maternal circulating levels and adiposity, indicating adiponectin is regulated within the mammary gland (27). Amongst its various functions, it has anti-inflammatory properties, breaks down fatty acids and heightens sensitivity to insulin. Strong correlations between adiponection levels in HM and infant serum (26) along with the presence of adiponectin receptors in the infant’s intestinal tract (26) emphasise its importance for the infant. Indeed, higher levels of circulating plasma adiponectin are related to lower BMI and healthier metabolism.

In animal models, adiponectin inhibits tension sensitive gastric vagal afferent mechanosensitivity, modulating satiety signals in both lean and obese animals, while simultaneously increasing the mechanosensitivity of

mucosal gastric vagal afferent in an obesity-induced model (28). In humans, elevated serum levels of adiponectin are associated with more rapid GE in diabetic patients (29). We are the first to have studied HM adiponectin in fully breastfed term infants and have found that increased levels and doses are associated with longer time between feeds in this population (16). This indicates adiponectin’s potential action on the gastric vagal nerve via decreased sensitivity of tension receptors, potentially resulting in delayed satiety signalling, leading to higher food intake.

The modulating effects of GE in infants and potentially on appetite and nutrient intake may partially explain the growth-regulating effect of adiponectin in infants in earlier months of life, as we and other researchers have shown (30). The relationship between changes in HM adiponectin and body composition and growth over the first two years of life has been studied. High levels of adiponectin are associated with lower weight, lean body mass and adiposity (30) (Gridneva and Geddes, unpublished data) in the first months of life, whereas in the second year of life, infants display more rapid growth and accretion of lean body mass and in a sense, ‘catch up’ to infants that received lower levels of adiponectin (31). Slower growth appears to be in part regulated by HM adiponectin and thus constitutes a potential mechanism for the lower risk of obesity in later life.

Ghrelin Ghrelin is a less studied orexigenic peptide produced

primarily in the stomach, but also in the lactating breast, resulting in HM having a higher ghrelin concentration than maternal serum (32). Ghrelin is an antagonist to leptin and stimulates appetite, gastric motility, acid secretion and food intake and is involved in long-term regulation of growth, weight and energy metabolism; thus heightened concentrations in HM could be vital for the infant drive to feed.

Ghrelin stimulates appetite and food intake and increases GE in rats (33) and humans (34) via the vagal nerve and afferent activity. High infant serum ghrelin levels are associated with increased age, length and weight in both formula-fed and breastfed infants (35,36), with formula-fed infants having higher ghrelin levels than breastfed infants (due to higher levels in formula) (10,35). Further, longer fasting times are associated with higher serum levels of ghrelin in formula-fed infants in the first six months of life (37). The inverse correlation between ghrelin and leptin in infant serum (10) indicates that HM leptin may play a role in lowering ghrelin levels. Since ghrelin and leptin actions are mediated by neuropeptide Y (NPY), reduced leptin levels allow ghrelin NPY stimulation and increase in food intake (10). Indeed, more comprehensive studies are required to clarify potential interactions between these appetite hormones.

HM ghrelin levels increase progressively in first months of lactation (38) and therefore, may influence infant feeding patterns and BC. Higher serum ghrelin levels in breastfed infants are associated with lower weight gain (36) and in formula-fed infants, with lower BMI (10), indicating a role for ghrelin in the regulation of body weight in healthy

SHOWCASE ON RESEARCHAppetite Control Hormones in Human Milk

Fig. 2. Measurements of infant body composition with bioimpedance spectroscopy.


SHOWCASE ON RESEARCH Appetite Control Hormones in Human Milk

infants. Higher ghrelin levels in HM are associated with higher infant weight gain (39,40) but lower weight (39). These somewhat contradictory results may be due to the combined effects of other HM hormones that were not measured in previous studies. Also it is not clear if ghrelin levels precede changes in appetite (37) and body weight (36,41) or follow them.

Ghrelin influences novelty seeking behaviour in rodents and humans (42), which may translate into an advantage in terms of locating new sources of food. Levels of ghrelin are blamed for impulsive shopping behaviour, as images of food become more desirable (43). These behaviour modulations mean that raised ghrelin levels in the infant will translate into hunger cues to gain the attention of the mother.

Other Key Appetite Hormones Insulin is a major hormone involved in glucose

metabolism and is produced by the pancreas. Studies in rats show that dietary insulin is not degraded in the stomach and plays an important role in the maturation and development of the small intestine (44); thus HM insulin could play a similar role in the development of human intestinal epithelium. Insulin is actively transported into HM in concentrations similar to maternal circulating levels (45) and is significantly higher (almost four times) than in bovine milk and infant formula, where it is barely detectable (46). While one study showed no differences in serum insulin concentration, fasting time and anthropometrics of breastfed and formula-fed infants (47), another study found that higher levels of milk insulin were related to lower weight and fat-free mass but not the fat mass in one month-old breastfed infants (19), reflecting insulin’s role as a regulator of energy balance, food intake and adiposity. Recently HM insulin has been associated with microbial diversity in the infant gut, with both insulin and leptin associated with beneficial microbial metabolic pathways that are able to reduce inflammation (48), further highlighting the diverse effects of these hormones.

Obestatin is an appetite-suppressing derivative of the ghrelin peptide precursor, which further emphasises the complexity of the appetite hormone system. Obestatin is produced by the cells lining the stomach and small intestine and salivary glands (49). Early studies reported that obestatin opposes ghrelin’s effects on body weight, food intake and GE (50). However, obestatin was later implicated in the inhibition of thirst and anxiety, regulation of sleep, memory improvement, induction of cell proliferation and increasing the secretion of the pancreatic juice enzymes (49). HM obestatin has been identified (51) with a concentration more than twice that of corresponding circulating maternal levels. Surprisingly however, the source of obestatin has still not been established. The relationship between obestatin and infant development and BC clearly requires further investigation.

A myriad of hormones have already been identified in HM and there are likely still many more to be discovered. Hormones such as apelin, resistin, insulin-like growth factor-I and motilin also play significant roles in the

metabolic development of infants and could be of interest in further research.

Conclusion

HM appetite hormones influence GE and BC in breastfed infants and are associated with early infant growth and development and subsequently health later in life. So the next time you see an infant being breastfed think, of all the delicate, complex mechanisms and pathways signalling to ensure programming of appetite regulation, leading to a reduced risk of obesity and chronic disease in our next generation.

References1. WHO (2007) Geneva, Switzerland2. Kent, J., Hepworth, A., Sherriff, J., Cox, D., Mitoulas, L.,

and Hartmann, P. (2013) Breastfeed. Med. 8, 401-4073. Butte, N., Wong, W., Hopkinson, J., Smith, E., and Ellis,

K. (2000) Pediatrics 16, 1355-13664. Goldman, A.S. (2000) J Nutr 130, 426S-431S5. Hassiotou, F., Savigni, D., Hartmann, P.E., and Geddes,

D.T. (2014) FASEB J. 28, 38.86. Kugananthan, S., Lai, C.T., Gridneva, Z., Mark, P.J.,

Geddes, D.T., and Kakulas, F. (2016) Nutrients 8, 7117. Savino, F., Benetti, S., Liguori, S., Sorrenti, M., and

Cordero Di Montezemolo, L. (2013) Cell. Mol. Biol. 59, 89-98

8. Pico, C., Oliver, P., Sanchez, J., Miralles, O., Caimari, A., Priego, T., and Palou, A. (2007) Int. J. Obes. 31, 1199-1209

9. Oliver, P., Pico, C., De Matteis, R., Cinti, S., and Palou, A. (2002) Dev. Dyn. 223, 148-154

10. Savino, F., Fissore, M.F., Grassino, E.C., Nanni, G.E., Oggero, R., and Silvestro, L. (2005) Acta Pediatr. 94, 531-537

11. Bouret, S.G., Draper, S.J., and Simerly, R.B. (2004) J. Neurosci. 24, 2797-2805

12. Proulx, K., Richard, D., and Walker, C. D. (2002) Endocrinology 143, 4683-4692

13. Smedh, U., Hakansson, M.L., Meister, B., and Uvnas-Moberg, K. (1998) Neuroreport 9, 297-301

14. Sanchez, J., Oliver, P., Miralles, O., Ceresi, E., Pico, C., and Palou, A. (2005) Endocrinology 146, 2575-2582

15. Cannon, A.M., Gridneva, Z., Hepworth, A., Lai, C.T., Tie, W.J., Khan, S., Hartmann, P.E., and Geddes, D.T. (2017) Pediatr. Res. in press

16. Gridneva, Z., Kugananthan, S., Hepworth, A.R., Tie, W.J., Lai, C.T., Ward, L.C., Hartmann, P.E., and Geddes, D.T. (2017) Nutrients 9, E15

17. Cannon, A., Kakulas, F., Hepworth, A., Lai, C., Hartmann, P., and Geddes, D. (2015) Int. J. Environ. Res. Public. Health 12, 12340-12355

18. Miralles, O., Sanchez, J., Palou, A., and Pico, C. (2006) Obesity 14, 1371-1377

19. Fields, D., and Demerath, E. (2012) Pediatr. Obes. 7, 304-312

20. Savino, F., Costamagna, M., Prino, A., Oggero, R., and Silvestro, L. (2002) Acta Pediatr. 91, 897-902

21. Savino, F., Liguori, S., Fissore, M., Palumeri, E.,


Calabrese, R., Oggero, R., Silvestro, L., and Miniero, R. (2008) J. Pediatr. Gastroenterol. Nutr. 46, 348-351

22. Martin, L., Woo, J., Geraghty, S., et al. (2006) Am. J. Clin. Nutr. 83, 1106-1111

23. Savino, F., Lupica, M., Benetti, S., Petrucci, E., Liguori, S., and Cordero Di Montezemolo, L. (2012) Acta Pediatr. 101, 1058-1062

24. Pajvani, U.B., Du, X., Combs, T.P., et al. (2003) J. Biol. Chem. 278, 9073-9085

25. Heinz, J.F.L. (2014) Adiponectin in cattle: profiling of molecular weight patterns in different body fluids at different physiological states and assessment of adiponectin’s effects on lymphocytes. PhD, Der Rheinischen Friedrich-Wilhelms-Universitat, Bonn

26. Newburg, D., Woo, J., and Morrow, A. (2010) J. Pediatr. 156, S41-S46

27. Weyermann, M., Beermann, C., Brenner, H., and Dietrich, R. (2006) Clin. Chem. 2, 2095-2102

28. Kentish, S.J., Ratcliff, K., Li, H., Wittert, G.A., and Page, A.J. (2015) Physiol. Behav. 152, 354-362

29. Iwase, M., Iino, K., Oku, M., Nohara, S., Asano, T., Doi, Y., and Iida, M. (2009) Diabetes Metab. Res. Rev. 25, 344-350

30. Woo, J., Guerrero, M., Altaye, M., Ruiz-Palacios, G., Martin, L., Dubert-Ferrandon, A., Newburg, D., and Morrow, A. (2009) Breastfeed. Med. 4, 101-109

31. Woo, J., Guerrero, M., Guo, F., Martin, L., Davidson, B., Ortega, H., Ruiz-Palacios, G., and Morrow, A. (2012) J. Pediatr. Gastroenterol. Nutr. 54, 532-539

32. Kierson, J., Dimatteo, D., Locke, R., Mackley, A., and Spear, M. (2006) Acta Pediatr. 95, 991-995

33. Asakawa, A., Inui, A., Kaga, T., et al. (2001) Gastroenterology 120, 337-345

34. Levin, F., Edholm, T., Schmidt, P.T., et al. (2006) J. Clin. Endocrinol. Metab. 91, 3296-3302

35. Savino, F., Petrucci, E., Lupica, M.M., Nanni, G.E., and Oggero, R. (2011) World J. Gastroenterol. 17, 1971-1975

36. Savino, F., Liguori, S., Fissore, M., Oggero, R., Silvestro, L., and Miniero, R. (2005) J. Pediatr. Gastroenterol. Nutr. 41, 653-659

37. Savino, F., Liguori, S., Grassino, E., Fissore, M., Guidi, C., Oggero, R., Silvestro, L., and Miniero, R. (2006) J. Pediatr. Gastroenterol. Nutr. 42, E86-E87

38. Ilcol, Y.O., and Hizli, B. (2007) Acta Paediatr. 96, 1632-1639

39. Kon, I.Y., Shilina, N.M., Gmoshinskaya, M.V., and Ivanushkina, T.A. (2014) Ann. Nutr. Metab. 65, 317-323

40. Khodabakhshi, A., Ghayour-Mobarhan, M., Rooki, H., et al. (2015) Eur. J. Clin. Nutr. 69, 614-618

41. Philipps, A.F. J. Pediatr. Gastroenterol. Nutr. 41, 575-57742. Hansson, C., Shirazi, R.H., Haslund, J., et al. (2012) PLoS

ONE 7, e5040943. Malik, S., McGlone, F., Bedrossian, D., and Dagher, A.

(2008) Cell Metab. 7, 400-40944. Buts, J.P., De Keyser, N., and Dive, C. (1988) Eur. J. Clin.

Nutr. 18, 391-39845. Whitmore, T., Trengove, N., Graham, D., and

Hartmann, P. (2012) Int. J. Endocrinol. 2012, ID 29636846. Shehadeh, N., Khaesh-Goldberg, E., Shamir, R.,

Periman, R., Sujov, P., Tamir, A., and Makhoul, I. (2003) Arch. Dis. Child. Fetal Neonatal Ed. 88, F214-F216

47. Savino, F., Grassino, E., Fissore, M., Guidi, C., Liguori, S., Silvestro, L., Oggero, R., and Miniero, R. (2006) Clin. Endocrinol. 65, 158-162

48. Lemas, D.J., Young, B.E., Baker, P.R., et al. (2016) Am. J. Clin. Nutr. 103, 1291-1300

49. Tang, S., Jiang, Q., Zhang, Y., et al. (2008) Peptides 29, 639-645

50. Zhang, J., Ren, P., Avsian-Kretchmer, O., Luo, C., Rauch, R., Klein, C., and Hsueh, A. (2005) Science 310, 996-999

51. Aydin, S., Ozkan, Y., Erman, F., et al. (2008) Nutrition 24, 689-693

SHOWCASE ON RESEARCHAppetite Control Hormones in Human Milk


The physiological processes of human lactation are not as foolproof as is often thought, as many women experience milk production difficulties, often leading to early weaning. Most of these difficulties are attributed to pain during feeding and real or perceived low milk supply. Some identified risk factors for impaired lactation include maternal obesity resulting in abnormal development of the mammary gland (1) and maternal mammary ZnT2 transporter mutations being linked to low production (2). Evidently, greater research into the mechanisms facilitating successful morphogenesis of the gland during pregnancy and lactation is desperately required to develop strategies to assist at-risk women and optimise their milk production. In the past, studies into the development of the gland have been hindered by the lack of readily accessible human lactating tissue (3). As a result, many previous studies have instead focused on animal models of pregnancy and lactation, although major differences exist in mammary gland development (4), gene expression (5) and milk composition (6) between mammalian species. However, human milk (HM) contains immune, stem and differentiated mammary

cells that can be used to non-invasively characterise lactating mammary cell gene expression, morphology and regenerative capabilities (Fig. 1).

Early studies examining cells from human colostrum (HC) and HM cells were conducted by smearing undiluted samples across slides to morphologically determine cell types and properties (7). Originally it was thought that milk was dominated by leukocytes and ‘foam cells’ (colostrum bodies) (7), however more modern techniques of cell isolation and classification have advanced our understanding of HM cells. Studies from our laboratory using freshly isolated cells extracted from either HC or HM examined for CD45 with flow cytometry found that whilst leukocytes dominate HC, they revert to <2.5% of total mature HM cells when both the mother and infant are healthy (8). On the other hand, infection of either the infant or mother can affect the leukocyte content of HM, increasing slightly for infant only infections and more dramatically (>90% leukocyte content) in cases of maternal mastitis (8,9) (Fig. 2). These findings, together with animal studies illustrating immune cell transfer through the milk to various infant organs (10),

particularly the thymus (11,12), suggests a potential protective and programming role of these cells for the infant. Additionally, the heightened levels of immune cells in cases of maternal mastitis provides a unique opportunity to study pathways of mammary gland defence that may be optimised for rapid resolution of mammary gland infection.

Unlike bovine milk, which contains low numbers of viable cells (primarily immune cells) (13,14), mature HM is a rich source of viable cells, representative of the lactating mammary epithelium (3,15) (Fig. 3A). Numerous viable cells are available for isolation from mature HM, ranging from 1.16x104 – 3.9x106 cells per millilitre of milk, of which 76-100% are viable (16-18). Lower numbers of viable cells can be isolated from either HC or early HM samples (18). Variation in these values are likely due to differences in isolation protocols, where storage and centrifugation speed are important in


Cells in Human Milk – What Do They Tell UsAlecia-Jane Twigger* and Peter Hartmann

School of Chemistry and Biochemistry, University of Western Australia, Perth, WA 6009*Corresponding author: [email protected]

Stem cell

Common progenitor

Myoepithelial progenitor

Myoepithelial Alveolar Ductal

Luminal progenitor

Figure 1

Fig. 1. Cells isolated from human milk contain a hierarchy of stem and differentiated mammary cells.


determining the number and viability of extracted cells (19). Recently, polyploidal luminal cells have been identified in the human mammary gland and these play a vital role in lactation, as observed by low milk production in mice with inhibited binucleated cell formation (20). It appears polyploidal cells filter into the milk as seen in Fig. 3A (Twigger, unpublished data), although more detailed imaging should be carried out to confirm this. Examination of HM cells has confirmed the expression of many key mammary markers including integrin alpha-6 (CD49f), membrane metalloendopeptidase (CD10), epithelial adhesion marker (EPCAM) and cytokeratins 5, 14, 18 and 19 (CK5, CK14, CK18 and CK19) which are representative of mammary stem and differentiated basal and luminal cells (16,21,22). Furthermore, a previous study found that mammary tissue and milk cells from lactating macaques had comparable gene expression, suggesting milk cells to be representative of mammary tissue (23). Discrepancies in HM cell gene expression have been associated with different characteristics of the mother and infant, building our understanding of the influencing factors that determine successful mammary gland development and milk production.

The population of cells in HM vary in composition over the course of a breastfeed as well as within and between women, much like milk composition in general (3). HM cell content varies over the course of a breastfeed, similar to the variation observed in fat content in HM, being greater towards the end of a feed and peaking approximately 30 minutes post-feeding (17). In a recent study, variation of HM cell gene expression was established between 66 participants for 17 different genes representative of the mammary cellular hierarchy (16). Gene expression was investigated in light of maternal and infant demographics, which included maternal body mass index (BMI), breast volume, gestational age of the infant and lactation stage, in order to determine the presence of any associations (16). Of note, lower expression of CK18, which is representative of lactocytes, was associated with higher maternal BMI, potentially suggesting undifferentiated lactocytes as an underlying cause of low milk production

in obese mothers (16). In addition, higher expression of estrogen related receptor beta (ESRRB) expression and lower expression of growth differentiation factor 3 (GDF3) expression were associated with earlier stages of lactation, implying ongoing development of the mammary gland over the course of lactation. Indeed whole transcriptome analysis between milk fat globules isolated from HC, transitional HM and mature HM found differences in expression of key genes such as milk protein genes alpha-lactatalbumin and beta casein between these three stages of lactation (24). Moreover, immune defence pathways were expressed specifically in HC, upregulation of milk synthesis machinery was found in transitional HM and lipid production pathways were dominant in mature HM (24). A subsequent study examined changes in the HM cell transcriptome by comparing sequential samples collected over the first 12 months of lactation with prepartum secretions from the same women and resting mammary gland tissue (25). As well as confirming findings from the HM fat globule study, this study determined differences in the expression of cell adhesion pathway genes between prepartum secretion cells and resting tissue, as well as differences in the expression of cancer pathway genes between mature HM cells and resting mammary tissue that were not identified in the HM fat globule study (25). Studies such as these will elucidate the mechanisms behind human milk synthesis and mammary metabolism that are trigged during secretory activation. It is evident that not only can HM cells be utilised to examine mammary gene expression (16,26,27), they can also be grown in culture to help further profile the developing and functioning mammary gland (21,22,26,28,29).

The first studies of HM cell culture were conducted in the late 1970s and early 1980s, where cells isolated from early milk (3 to 7 days post-partum) proliferated in 2D culture and survived multiple passages (30-33). Interestingly, it was noted that not all cells participated in cell division whilst many of the cells could proliferate for 8–11 passages (30). Subsequent studies found cells isolated from more mature HM could also be cultured and these displayed many of the features originally

SHOWCASE ON RESEARCHCells in Human Milk

Forward Scatter

Side

Sca

tter

A

i

iii ii

CD45

55% CD45+

CD45

70.5% CD45+

B i) B ii)

Cou

nt

Fig. 2. Flow cytometric analysis of isolated milk cells collected from a woman with mastitis.A. Forward and side scatter plots reveal size and granularity of milk cells which were gated to examine subpopulations i), ii) and iii).B. Cells from subpopulations i) and ii) were examined for CD45, where subpopulation i) had 70.5% of cells positive for CD45, whereas subpopulation ii) had 55% of cells positive for CD45.


SHOWCASE ON RESEARCH Cells in Human Milk

Fig. 2. Human milk cells examined with an inverted microscope.A. Freshly isolated cells stained with Trypan Blue exclusion dye illustrate a large number of viable cells with some apparent bi-nucleated cells (red arrows), non-viable cells (black arrow) and few milk fat globules (yellow arrow).B. Cells placed in mammary media for two days begin to forming spheroids under non-adherent conditions.

described with added immunofluorescence staining, demonstrating the diversity in the resulting mammary cell subtypes (21,22,26,28). Additionally, it was found that CK5 and CK14 gene expression was enriched after culture, providing an opportunity to select and expand the progenitor cell population from freshly isolated HM cells (21). Another study using cultured monolayer HM cells was able to identify differences in the morphology and growth patterns of the cells after transfection of 14-3-3s (Sigma) siRNA (28). Similar gene silencing techniques could be utilised in the future to examine the importance of specific genes for mammary gland development. Using non-adherent culturing conditions, 3D structures can also be formed from HM cells (Fig. 3B) to create bi-layered spheroids containing CD49f+ positive basal cells and Prl-R+ (prolactin receptor) luminal cells, mimicking mammary alveoli (22). These 3D structures were shown to synthesise and secrete the milk proteins alpha-lactalbumin and lactoferrin into the culture media (26). Enhancements of these protocols may utilise the latest organoid culture methods, which are able to produce organised tertiary alveolar and conn3cting ductal structures from resting mammary cells (34). Indeed, HM cell culture may hold the key to a greater understanding of human mammary gland development, which is driven by progenitor and stem cells.

Currently, human mammary stem cells are not well defined with respect to markers, location or differentiation capabilities due to the limited availability of tissue and heterogeneous methods of investigation (35). It is likely that a spectrum of mammary stem cells exists with different differentiation capabilities. Investigations of lactating mammary cells isolated through HM have found subpopulations that are positive for markers associated with bipotent, multipotent and pluripotent markers (21,26,28). Nestin was the first multipotent stem cell marker identified in HM cells and co-stained with the mammary stem cell marker CK5 (21). HM cells positive for multipotent marker p63 have also been identified and

can differentiate into two separate mammary cell lineages and consequently undergo cell cycle arrest (28). Lineage-specific progenitor markers such as CD105, CD29, CD90 and Stro-1, which are representative of mesenchymal stem cells, have been identified in HM, which are able to secrete vascular endothelial growth factor (VEGF) in culture (36-39). Whether these cells are true mesenchymal stem cells is debatable (40), however further studies suggest that HM cells exhibit lineage plasticity. An interesting study by Hassiotou et al. demonstrated the presence of pluripotent transcription factors OCT4, SOX2 and NANOG in HM cells and illustrated their plasticity by differentiating them into neuronal, hepatocyte, pancreatic and bone–like cells (26). Similar studies have been conducted in resting mammary tissue, which also contain highly plastic cells (41), albeit at lower densities (26). Evidently, stem cells in the mammary gland are vital to achieving maturity in pregnancy and to some degree lactation. However it is not known why these cells are excreted into the milk. It is possible that they exist for the benefit of the infant, where they have been found to survive the gastrointestinal tract of the offspring and are transferred to distant tissues where they integrate to potentially contribute to tissue function (42). It has been speculated that HM cells have a potential use in regenerative therapies; however a great deal more work is needed to understand the cells’ characteristics and proportions in milk.

Clearly HM contains a fascinating cellular composition that can be non-invasively isolated and used in a plethora of experiments to examine lactating mammary cell gene expression, growth and in vitro morphology. HM cells will also assist in the understanding of the intricacies of mammary gland development and function. Future milk cell work investigating the characteristics of normal HM cells hold the promise of unravelling mammary gland dysfunction and pathologies such as low milk production and mastitis, both of which are major contributors to early weaning.


References1. Rasmussen, K.M. (2007) Ann. Rev. Nutr. 27, 103-1212. Dempsey, C., McCormick, N.H., Croxford, T.P., Seo,

Y.A., Grider, A., and Kelleher, S.L. (2012) J. Nutr. 142, 655-660

3. Hassiotou, F., Geddes, D.T., and Hartmann, P.E. (2013) J. Hum. Lact. 29, 171-182

4. Dontu, G., and Ince, T.A. (2015) J. Mammary Gland Biol. Neoplasia 20, 51-62

5. Lemay, D.G., Lynn, D.J., Martin, W.F., Neville, M.C., Casey, T.M., Rincon, G., Kriventseva, E.V., Barris, W.C., Hinrichs, A.S., Molenaar, A.J., Pollard, K.S., Maqbool, N.J., Singh, K., Murney, R., Zdobnov, E.M., Tellam, R.L., Medrano, J.F., German, J.B., and Rijnkels, M. (2009) Genome Biol. 10, R43

6. Malacarne, M., Martuzzi, F., Summer, A., and Mariani, P. (2002) Int. Dairy J. 12, 869-877

7. Holmquist, D.G., and Papanicolaou, G.N. (1956) Ann. NY Acad. Sci. 63, 1422-1435

8. Hassiotou, F., Hepworth, A.R., Metzger, P., Lai, C.T., Trengove, N., and Hartmann, P.E. (2013) Clin. Transl. Immunology 2, e3

9. Riskin, A., Almog, M., Peri, R., Halasz, K., Srugo, I., and Kessel, A. (2012) Pediatr. Res. 71, 220-225

10. Jain, L., Vidyasagar, D., Xanthou, M., Ghai, V., Shimada, S., and Blend, M. (1989) Arch. Dis. Child 64, 930-933

11. Zhou, L., Yoshimura, Y., Huang, Y., Suzuki, R., Yokoyama, M., Okabe, M., and Shimamura, M. (2000) Immunology 101, 570-581

12. Ghosh, M.K., Nguyen, V., Muller, H.K., and Walker, A.M. (2016) J. Immunol. 197, 2290-2296

13. Lee, C.-S., Wooding, P., and Kemp, P. (1980) J. Dairy Res. 47, 39-50

14. Boutinaud, M., and Jammes, H. (2002) Reprod. Nutr. Dev. 42, 133-147

15. Ho, F.C., Wong, R.L., and Lawton, J.W. (1979) Acta Paediatr. Scand. 68, 389-396

16. Twigger, A.J., Hepworth, A.R., Lai, C.T., Chetwynd, E., Stuebe, A.M., Blancafort, P., Hartmann, P.E., Geddes, D.T., and Kakulas, F. (2015) Sci. Rep. 5, 12933

17. Hassiotou, F., Hepworth, A.R., Williams, T.M., Twigger, A.J., Perrella, S., Lai, C.T., Filgueira, L., Geddes, D.T., and Hartmann, P.E. (2013) PLoS One 8, e78232

18. Gaffney, E.V., Polanowski, F.P., Blackburn, S.E., and Lambiase, J.P. (1976) Cell Tissue Res. 172, 269-279

19. Hassiotou, F., Savigni, D.L., Hartmann, P., and Geddes, D.T. (2015) FASEB J. 29

20. Rios, A.C., Fu, N.Y., Jamieson, P.R., Pal, B., Whitehead, L., Nicholas, K.R., Lindeman, G.J., and Visvader, J.E. (2016) Nat. Commun. 7, 11400

21. Cregan, M.D., Fan, Y., Appelbee, A., Brown, M.L., Klopcic, B., Koppen, J., Mitoulas, L.R., Piper, K.M., Choolani, M.A., Chong, Y.S., and Hartmann, P.E. (2007) Cell Tissue Res. 329, 129-136

22. Thomas, E., Lee-Pullen, T., Rigby, P., Hartmann, P., Xu, J., and Zeps, N. (2012) Stem Cells 30, 1255-1264

23. Lemay, D.G., Hovey, R.C., Hartono, S.R., Hinde, K., Smilowitz, J.T., Ventimiglia, F., Schmidt, K.A., Lee, J., Islas-Trejo, P., Silva, P.I., Korf, I., Medrano, J., Barry, P.A., and German, J.B. (2013) BMC Genomics 14, 872

24. Lemay, D.G., Ballard, O.A., Hughes, M.A., Morrow, A.L., Horseman, N.D., and Nommsen-Rivers, L.A. (2013) PLoS One 8, e67531

25. Twigger, A.J., Geddes, D.T., and Kakulas, F. (2016) 18th International Society for Research into Human Milk and Lactation Conference, Breastfeeding Medicine, Stellenbosch

26. Hassiotou, F., Beltran, A., Chetwynd, E., Stuebe, A.M., Twigger, A.J., Metzger, P., Trengove, N., Lai, C.T., Filgueira, L., Blancafort, P., and Hartmann, P.E. (2012) Stem Cells 30, 2164-2174

27. Sharp, J.A., Lefevre, C., Watt, A., and Nicholas, K.R. (2016) Funct. Integr. Genomics 16, 297-321

28. Thomas, E., Zeps, N., Cregan, M., Hartmann, P., and Martin, T. (2011) Cell Cycle 10, 1-7

29. Hosseini, S.M., Talaei-Khozani, T., Sani, M., and Owrangi, B. (2014) Neurol. Res. Int. 2014, 807896

30. Chang, S.E., Keen, J., Lane, E.B., and Taylor-Papadimitriou, J. (1982) Cancer Res. 42, 2040-2053

31. Stoker, M., Perryman, M., and Eeles, R. (1982) Proc. R. Soc. Lond. B. Biol. Sci. 215, 231-240

32. Gaffney, E.V. (1982) Cell Tissue Res. 227, 563-56833. Taylor-Papadimitriou, J., Shearer, M., and Tilly, R.

(1977) J. Natl. Cancer Inst. 58, 156334. Linnemann, J.R., Miura, H., Meixner, L.K., Irmler, M.,

Kloos, U.J., Hirschi, B., Bartsch, H.S., Sass, S., Beckers, J., Theis, F.J., Gabka, C., Sotlar, K., and Scheel, C.H. (2015) Development 142, 3239-3251

35. Visvader, J.E., and Stingl, J. (2014) Genes Dev. 28, 1143-1158

36. Fan, Y., Chong, Y.S., Choolani, M.A., Cregan, M.D., and Chan, J.K. (2010) PLoS One 5, e14421

37. Indumathi, S., Dhanasekaran, M., Rajkumar, J. S., and Sudarsanam, D. (2012) Cytotechnology 65, 385-393

38. Patki, S., Kadam, S., Chandra, V., and Bhonde, R. (2010) Hum. Cell 23, 35-40

39. Kaingade, P.M., Somasundaram, I., Nikam, A.B., Sarang, S.A., and Patel, J.S. (2016) Breastfeed. Med. 11, 26-31

40. Kakulas, F., Geddes, D. T., and Hartmann, P. E. (2016) Breastfeed. Med. 11, 150-151

41. Roy, S., Gascard, P., Dumont, N., Zhao, J., Pan, D., Petrie, S., Margeta, M., and Tisty, T. (2013) Proc. Natl. Acad. Sci. USA 110, 4598-4603

42. Hassiotou, F., Mobley, A., Ocal, O., Filgueira, L., Geddes, D.T., Hartmann, P.E., and Wilkie, T.M. (2014) 17th Conference of International Society for Research in Human Milk and Lactation

SHOWCASE ON RESEARCHCells in Human Milk


Lactation has evolved over the past 200 million years since the appearance of the aplacental, egg laying monotremes. Subsequently, there has been extensive adaptation to reproduction, particularly in lactational strategies, when the Theria (subclass of mammals) split into the Metatheria (Marsupialia) and Eutheria (Placentalia) lineages over 140 million years ago (1). The role of milk in providing nutrition to the suckled young in mammals is well established. However, over the past few decades a focus on the functionality of milk has confirmed that it also delivers a range of bioactives that are either present in milk in the mammary gland or are released when the milk is processed in the oral cavity and the gut of the neonate. These bioactives may include proteins, peptides, complex oligosaccharides and miRNA and have the potential to stimulate development, regulate gut flora and provide protection for the young from infection (2-8). An interesting and often neglected group of bioactives is now being identified; they have a function locally within the mammary gland to remodel the mammary tissue (stimulate growth or signal cell death) and can provide protection from infection and inflammation when the mammary gland is susceptible to these challenges. This review will focus predominantly on the relationship between milk proteins and proteases, resulting in the release of bioactive peptides.

Milk Bioactives – Impact on Development of the YoungTo better identify bioactives, scientists have increasingly

used comparative genomics and bioinformatics to explore the lactational strategies of the Australian monotremes and marsupials. The lactation cycle in the only two extant monotremes, the echidna (Tachyglossus and Zaglosus genera) and platypus (Ornithorhynchus anatinus), and an Australian marsupial, the tammar wallaby (Macropus eugenii) have been studied extensively (9,10). The echidna lays shelled eggs (11) and the hatchlings are altricial (undeveloped) and not immune competent (12). Therefore, milk is not only important for growth and development but particularly for protection of the young from disease while they are in a non-sterile environment (13,14).

Reproduction in the tammar is characterised by a short gestation period (26.5 days), birth of immature young and a long lactation period (approximately 300 days), during which the concentration of all the major milk constituents, and many minor milk bioactives progressively change (15) (Fig. 1A). There is increasing evidence that these changes in milk composition regulate growth of the tammar pouch young (10,16), particularly during the first 100 days postpartum, when the development of the neonate is similar to a late stage eutherian fetus. Therefore the signalling factors involved in the development of the eutherian fetus are most likely delivered to marsupials in the milk of the mother (17) (Fig. 1B). Fostering experiments demonstrated that transferring the early phase pouch young to a late phase lactating tammar can accelerate the growth and physical development of pouch young (18,19), and also accelerate maturation of specific organs such as the stomach (20). More recent studies using in vitro models have shown that milk collected from marsupials during early lactation (day 20–100), but not late lactation (day 100–300), stimulated proliferation and differentiation of whole lung cultures from mouse embryos (21). Therefore the temporal delivery of these bioactives is most likely crucial for the development of the suckled young.

The regulation of the lactation cycle in the tammar wallaby has fascinated and challenged scientists for many decades and the interesting interplay between the endocrine, autocrine and paracrine mechanisms that are implicated in this process are now beginning to be better understood. If these animals are genuine biomedical research models that offer new insights into reproduction in eutherians, it must be assumed that ‘evolution has not reinvented the wheel’ and that the basic mechanisms regulating cellular responses have been conserved over the past 140 million years.

This assumption appears to be reasonably sound. Research over the past 50 years has shown that insulin, glucocorticoids and prolactin are required for lactation in eutherians and a number of in vitro studies have shown these hormones are a basic requirement for successful lactation in the tammar and echidna (22-26). It is now accepted that the mammary extracellular matrix (ECM) has


Human Milk Bioactivity:Lessons from the Evolution of Lactation

Julie Sharp1,2*, Ashalyn Watt1, Christophe Lefevre3,4,5 and Kevin Nicholas6,7

1Institute for Frontier Materials, Deakin University, Geelong, VIC 32162Cancer Program, Monash Biomedicine Discovery Institute and Department ofAnatomy and Developmental Biology, Monash University, Clayton, VIC 3800

3Division of Bioinformatics, Walter and Eliza Hall Medical Research Institute, Melbourne, VIC 30004Peter MacCallum Cancer Centre, Melbourne, VIC 3000

5Department of Medical Biology, University of Melbourne, Melbourne, VIC 30006Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800;

7School of Biosciences, University of Melbourne, Melbourne, VIC 3010*Corresponding author: [email protected]


an integral role and is essential for the development of the mammary gland during gestation, the overt differentiation of mammary epithelial cells for galactopoesis and remodelling of the mammary gland during involution (27-29). More recent studies have shown that the ECM in the tammar has additional roles in regulating the cellular program for the lactation cycle in the tammar, particularly in controlling the progressive changes in milk composition (30,31). Perhaps one of the most exciting and compelling studies on the regulation of lactation was reported recently by Jane Visvader’s laboratory (29). The experiments using a mouse model showed that binuclear mammary epithelial cells appeared in the mammary gland at lactogenesis and disappeared at involution. The presence of these cells in the mammary gland was essential for successful lactation. However it was fascinating to note that these binuclear cells were also present in lactating tissue from the human and wallaby, indicating that this basic regulatory process was conserved during the evolution of lactation.

It is ironic that marsupials may be considered a primitive

mammal. However, the reality is that the mammary gland in these species is very sophisticated in terms of its capacity for temporal delivery of bioactives for multiple targets. The eutherian mammary gland is less sophisticated as many of its previous functions have evolved to be delivered by multiple tissues, particularly a well-developed placenta and the amniotic fluid. It is clear that the marsupial provides a unique opportunity to more easily identify bioactives that potentially play a role in the early development of the fetus. However, it may suggest that an increased focus on human colostrum would be appropriate to get a better understanding of its potential role in regulating early development of the baby, particularly at a time when the gut is amenable to increased transfer of milk components.

We have known for some time that significantly premature and low birthweight babies have acute challenges for survival, largely due to limited development of their lungs and gut, and these babies may show an increased frequency of mature onset disease (32). Studies using the tammar wallaby may lead to a new range of

Fig. 1. The lactation cycle of the tammar wallaby.A. The lactation cycle in the tammar has been divided into four phases, characterised by changes in milk composition and the sucking pattern of the pouch young.B. Development of the tammar pouch young from day 6 to day 220 postpartum (upper panels) compared to development of the human embryo at day 23 and preterm baby at 24 weeks of age (lower panels).


Human Milk Bioactivity


SHOWCASE ON RESEARCH Human Milk Bioactivity

human fortifiers that include bioactives with the potential to specifically target the growth and development of tissues in the human neonate to improve outcomes for premature and low birthweight babies (33,34).

Therefore, developmental clocks are set in the neonate and any potential disruption to this process may subsequently impact on mature onset disease. The tammar provides a new model to better understand this process of developmental programming. It is not known if this developmental program is set during the short gestation or whether the milk provided in early lactation includes signals to the altricial neonate that have a role in this process. The option of cross fostering neonates to mothers at advanced stages of lactation to exclude the delivery of putative milk bioactives to the suckled young (33-35) may shed new light on the initiation of developmental programming.

Milk Bioactives That Impact on Mammary Function The early work of Li et al. (36) showed that ligating

the teats on one side of a lactating mouse led to these mammary glands progressing only to involution, despite all mammary glands being exposed to the same hormonal milieu. This was clear evidence that milk bioactivity could impact locally on mammary gland physiology. More recent work using the tammar has shown that the mammary gland has a sophisticated capacity to use milk when under challenge from infection and that milk can stimulate growth, and program cell death of the tissue when appropriate (36,37). Mammary genes coding for milk proteins can be multifunctional and demonstrate a temporal domain-specific bioactivity resulting from alternate splicing (38). For example, cathelicidin usually exists in an inactive proform and is cleaved by specific proteases to provide the two domains of cathelicidin that may have a variety of functions (39). The tammar cathelicidin 1 gene (MaeuCath1) is differentially expressed in the mammary gland throughout the lactation cycle as two splice variants (MaeuCathel 1a and 1b) (38). The level of MaeuCath1a transcripts are upregulated only during early lactation (2A) and late involution whereas MaeuCath1b transcripts are expressed throughout lactation and steadily increase from late lactation until early involution. The protein corresponding to MaeuCath1a significantly inhibits a range of bacteria (38). The expression of this transcript in the first 48 hours postpartum to day 80 postpartum is consistent with a need to provide protection from pathogens (40). MaeuCath1a expression at day 10 of involution suggests an additional antibacterial role at a time when the mammary gland is more susceptible to pathogen-mediated mastitis (41). The continued expression of the MaeuCath1b transcript when the neonate has developed adaptive immunity suggests additional roles for the maintenance and proliferation of mammary epithelia during increasing milk production (23,42). Indeed, several studies have suggested a role for cathelicidins in epithelial cell proliferation during wound healing, maintenance and re-establishment of the intestinal barrier integrity and proliferation of lung epithelial cells (43-45). More recent in vitro studies have shown

wallaby mammary epithelial cells exhibiting increased proliferation following inclusion of MaeuCath1b in the media, confirming this hypothesis (38).

Another interesting tammar milk protein, WAP four-disulphide domain protein-2 (WFDC2) is part of a large family of whey acidic protein (WAP) four disulfide core proteins (46-49). Tammar WFDC2 is comprised of two four-disulfide core domains annotated as domain III on the amino terminal end and domain II at the carboxyl terminal end (50). Expression of the WFDC2 gene is elevated only in pregnancy, early lactation and during involution (51). Studies by Watt et al. (52) showed domain II of the protein had antibacterial activity against a range of pathogenic bacteria but no antibacterial activity against Enterococcus faecalis. The elevated expression of WFDC2 during pregnancy and involution correlates with the timing of increased risk of infection in the mammary gland (40,52,53), largely resulting from Staphylococcus aureus, Streptococcus spp. and Escherichia coli in the mammary tissue (54-56). However, expression of this gene during the first 100 days postpartum suggests an additional role in protecting the pouch young when it is not immune competent (52). Therefore, the antibacterial effect of WFDC2 appears to be directed to pathogenic bacteria and not commensal bacteria (40,53,57). The down regulation of WFDC2 around 100 days postpartum, when the young detaches from the teat, correlates with the development of an immune response in the young.

Human Milk Bioactivity – Acute Response to ChallengeThe examples of the temporal regulation of either intact

proteins or domain-specific expression of proteins to provide the kind of milk bioactivity required by the tammar mammary gland and the suckled young is dependent on sophisticated programming of the mammary gland. However, as discussed previously, there is a reduced need for this level of progressive change in the delivery of specific bioactives in human milk, although it is evident that human milk composition can change during lactation and in response to specific challenges to the breast (58,59).

A recent study by Sharp et al. (2016) using microarrays to analyse the expression of genes in cells present in breast milk at days 24, 48 and 101 of lactation and days 7 and 14 of involution identified a total of 206 protease genes (60). A large component of these genes were predicted to code for proteins with a signal peptide. Predicted classes of secreted proteases included threonine, aspartate, cysteine, serine and metallo proteases. Some of these genes were expressed throughout lactation whereas other genes were expressed only at specific time points during lactation. Bioactive milk peptides resulting from proteolytic digestion had functions that included immunomodulatory (61,62), antimicrobial (63-65), and antithrombotic (66,67) activity, and these peptides could act as opioid agonists (68-71), ACE inhibitors (72) and proliferative factors (73).

In recent studies, skim breast milk was incubated at 37 degrees over a period of 5 days to allow digestion of milk protein by endogenous proteases (Watt, Nicholas and Sharp, unpublished data). Peptides were predominantly derived from casein proteins, and antibacterial assays




using mastitis causing S. aureus showed that incubating this hydrolysate significantly increased antimicrobial activity. Similarly, these peptides showed anti-inflammatory activity. Importantly, the peptides did not show any capacity to program apoptotic activity in human mammary epithelial cells. Therefore these peptides may play a specific role in the breast to reduce infection and inflammation if there is an interruption to breastfeeding.

Acute Responses to Mastitic ChallengeDetermination of gene expression in normal lactating

women demonstrated the utility of RNA obtained directly from human milk cells to detect mammary epithelial cell (MEC)-specific gene expression. Therefore milk cell RNA collected during mastitis was also analysed by human Affymetrix arrays and revealed regulation of a unique set of genes specific to this disease state, whilst maintaining regulation of milk synthesis genes (60). Genes specific to infection and related to immune function, antimicrobial response, and anti-inflammatory response were upregulated compared to healthy milk (Fig. 2). More recent studies (Watt, Nicholas and Sharp, unpublished data) have used human MECs cultured on a range of ECMs which allows the formation of three-dimensional acini (mammospheres) that respond to insulin, cortisol and prolactin to express milk protein genes. These mammospheres were challenged with lipoteichoic acid (LTA) and showed similar transcriptional responses to milk-derived cells from women with mastitis (Fig. 3). These in vitro acini therefore represent a robust and relevant human mammary model to better understand the acute molecular responses to mastitic challenge, and may therefore provide new opportunities for improved treatment of breast infection.

This review has focused on the relationship between milk proteins and peptide bioactivity. Although it is becoming evident that breastmilk bioactivity for the development of the young has been diminishing with a greater contribution from the placenta, there is still much

to explore with regard to the complexity of human milk. The established role of complex carbohydrate on gut flora and the general health of the young and the emerging role of miRNA as legitimate signalling molecules for both the breast and the young (74,75) emphasise the complexity of the multiple roles of milk, in addition to its role in nutrition. However, it is increasingly evident that the interrogation of the evolutionary history of lactation has great promise for the application of comparative approaches to obtain a better understanding the role of milk in acute and chronic wellbeing of the baby.

References1. Lefevre, C.M., Sharp, J.A., and Nicholas, K.R. (2010)

Annu. Rev. Genomics Hum. Genet. 11, 219-2382. FitzGerald, R.J., Murray, B.A., and Walsh, D.J. (2004) J.

Nutr. 134, 980S-988S3. Hartmann, R. and Meisel, H. (2007) Curr. Opin.

Biotechnol. 18, 163-1694. Allen-Blevins, C.R., Sela, D.A., and Hinde, K. (2015) Evol.

Med. Public Health 2015, 106-1215. Zivkovic, A.M., German, J.B., Lebrilla, C.B., and Mills,

D.A. (2011) Proc. Natl. Acad. Sci. USA 108 Suppl 1, 4653-4658

6. Marcobal, A., Barboza, M., Froehlich, J.W., et al. (2010) J. Agric. Food Chem. 58, 5334-5340

7. Zhou, Q., Li, M., Wang, X., et al. (2012) Int. J. Biol. Sci. 8, 118-123

8. Admyre, C., Johansson, S.M., Qazi, K.R., et al. (2007) J. Immunol. 179, 1969-1978

9. Sharp, J.A., Watt, A., Bisana, S., et al. In: Singh, H., Thompson, A., and Boland, M. (eds.), Milk Proteins: From Expression to Food (2014) Elsevier, Chapter 3, pp76-112

10. Sharp, J.A., Modepalli, V., Enjapoori, A.K., et al. (2015) J. Mammary Gland Biol. Neoplasia 19, 289-302

11. Morrow, G.E., and Nicol, S.C. (2013) Aust. J. Zool. 60, 289-298

12. Griffiths, M. The Biology of Monotremes (1978) Acedemic Press, USA

Fig. 2. Mastitis-specific gene expression profile. Expression profiles of genes related to a defence response within cells collected from healthy milk and mastitic milk. Y axes represent expression values. COL: colostrum 24 hours prepartum; LACT: lactation at day 24, 48, 101; MAST; mastitis at day 23; INV: involution at day 7 and 14.


SHOWCASE ON RESEARCH Human Milk Bioactivity

13. Bisana, S., Kumar, S., Rismiller, P., et al. (2013) PLoS One 8, e53686

14. Enjapoori, A.K., Grant, T.R., Nicol, S.C., et al. (2014) Genome Biol. Evol. 6, 2754-73

15. Tyndale-Biscoe, C.H., and Janssens, P.A. The Developing Marsupial: Models for Biomedical Research (1988) Springer-Verlag, Germany

16. Sharp, J.A., Lefevre, C., Kwek, J., et al. In: Deakin, J.E., Waters, P.D., and Marshall Graves, J.A. (eds.), Marsupial Genetics and Genomics (2010) Springer, UK. pp317-334

17. Brennan, A.J., Sharp, J.A., Digby, M.R., and Nicholas, K.R. (2007) IUBMB Life 59, 146-150

18. Menzies, B.R., Shaw, G., Fletcher, T.P., and Renfree, M.B. (2007) Reprod. Fert. Develop. 19, 976-983

19. Trott, J.F., Simpson, K.J., Moyle, R.L., et al. (2003) Biol. Reprod. 68, 929-36

20. Kwek, J., De Iongh, R., Nicholas, K., and Familari, M. (2009) J. Exp. Zool. B Mol. Dev. Evol. 312, 613-24

21. Modepalli, V., Hinds, L.A., Sharp, J.A., Lefevre, C., and Nicholas, K.R. (2015) BMC Dev. Biol. 15, 16

22. Nicholas, K.R. and Tyndale-Biscoe, C.H. (1985) J. Endocrinol. 106, 337-42

23. Bird, P.H., Hendry, K.A., Shaw, D.C., Wilde, C.J., and Nicholas, K.R. (1994) J. Mol. Endocrinol. 13, 117-25

24. Ballard, F.J., Grbovac, S., Nicholas, K.R., Owens, P.C., and Read, L.C. (1995) Gen. Comp. Endocrinol. 98, 262-8

25. Kwek, J.H., Wijesundera, C., Digby, M.R., and Nicholas, K.R. (2007) Biochim. Biophys. Acta 1770, 48-54

26. Enjapoori, A.K., Lefevre, C.M., Nicholas, K.R., and Sharp, J.A. (2017) Gen. Comp. Endocrinol. 242, 38-48

27. Roskelley, C.D., Srebrow, A., and Bissell, M.J. (1995) Curr. Opin. Cell Biol. 7, 736-47

28. Schedin, P., Mitrenga, T., McDaniel, S., and Kaeck, M. (2004) Mol. Carcinog. 41, 207-20

29. Rios, A.C., Fu, N.Y., Jamieson, P.R., et al. (2016) Nat. Commun. 7, 11400

30. Wanyonyi, S.S., Lefevre, C., Sharp, J.A., and Nicholas, K.R. (2013) Matrix Biol. 32, 342-51

31. Wanyonyi, S.S., Lefevre, C., Sharp, J.A., and Nicholas, K.R. (2013) Dev. Comp. Immunol. 40, 289-99

Fig. 3. Mastitis-specific gene expression mimicked by in vitro mammospheres. Expression profiles of genes which were shown to be upregulated in mastitis were also induced when mammospheres were incubated with LTA for 4 and 24 hours.A. Haematoxylin and eosin stain of paraffin embedded milk cells collected from mature milk. Arrows indicate presence of intact cells showing stained nuclei and cytoplasm. Binucleated cells can be observed.B–D. Gene expression profile of milk cells collected from the different stages of lactation (24, 48 and 101 days) and mastitis (day 23).E. Bright-field microscopy of human mammospheres grown for 7 days.F–H. Gene expression profiles of 7 day mammospheres treated with and without LTA for 4 and 24 hours.




32. Svedenkrans, J., Henckel, E., Kowalski, J., Norman, M., and Bohlin, K. (2013) PLoS One 8, e80869

33. Waite, R., Giraud, A., Old, J., et al. (2005) J. Exp. Zool. A Comp. Exp. Biol. 303, 331-344

34. Kwek, J.H., Iongh, R.D., Digby, M.R., et al. (2009) Mech. Dev. 126, 449-463

35. Modepalli, V., Hinds, L.A., Sharp, J.A., Lefevre, C., and Nicholas, K.R. (2016) Mech. Dev. 142, 22-29

36. Khalil, E., Digby, M.R., O’Donnell, P., and Nicholas, K.R. (2008) Comp. Biochem. Physiol. B Biochem. Mol. Biol. 151, 64-69

37. Brennan, A.J., Sharp, J.A., Lefevre, C.M., and Nicholas, K.R. (2008) J. Mol. Endocrinol. 41, 103-116

38. Wanyonyi, S.S., Sharp, J.A., Khalil, E., Lefevre, C., and Nicholas, K.R. (2011) Comp. Biochem. Physiol. A Mol. Integr. Physiol. 160, 431-439

39. Yang, D., Biragyn, A., Hoover, D.M., Lubkowski, J., and Oppenheim, J.J. (2004) Annu. Rev. Immunol. 22, 181-215

40. Daly, K.A., Digby, M., Lefevre, C., et al. (2007) Vet. Immunol. Immunopathol. 120, 187-200

41. Oliver, S.P., and Mitchell, B.A. (1983) J. Dairy Sci. 66, 1162-1166

42. Dove, H., and Cork, S.J. (1989) J. Zool. 219, 385-39743. Heilborn, J.D., Nilsson, M.F., Kratz, G., et al. (2003) J.

Invest. Dermatol. 120, 379-38944. Otte, J.M., Zdebik, A.E., Brand, S., et al. (2009) Regul. Pept.

156, 104-11745. Shaykhiev, R., Beisswenger, C., Kandler, K., et al. (2005)

Am. J. Physiol. Lung Cell Mol. Physiol. 289, L842-L84846. Bingle, L., Singleton, V., and Bingle, C.D. (2002) Oncogene

21, 2768-277347. Campbell, S.M., Rosen, J.M., Hennighausen, L.G.,

Strech-Jurk, U., and Sippel, A.E. (1984) Nucleic Acids Res. 12, 8685-8697

48. Topcic, D., Auguste, A., De Leo, A.A., et al. (2009) Evol. Dev. 11, 363-375

49. Hennighausen, L.G., and Sippel, A.E. (1982) Nucleic Acids Res. 10, 2677-2684

50. Sharp, J.A., Lefevre, C., and Nicholas, K.R. (2007) Evol. Dev. 9, 378-392

51. Watt, A.P., Sharp, J.A., Lefevre, C., and Nicholas, K.R. (2012) Dev. Comp. Immunol. 36, 584-590

52. Basden, K., Cooper, D.W., and Deane, E.M. (1997) Reprod. Fertil. Dev. 9, 243-254

53. Old, J.M., and Deane, E.M. (2000) Dev. Comp. Immunol. 24, 445-454

54. Barkema, H., Green, M., Bradley, A., and Zadoks, R. (2009) J. Dairy Sci. 92, 4717-4729

55. Borm, A., Fox, L., Leslie, K., et al. (2006) J. Dairy Sci. 89, 2090-2098

56. Bradley, A.J., and Green, M.J. (2001) J. Clin. Microbiol. 39, 1845-1849

57. Old, J.M., and Deane, E.M. (1998) Comp. Immunol. Microbiol. Infect. Dis. 21, 237-245

58. Ballard, O., and Morrow, A.L. (2013) Pediatr. Clin. North Am. 60, 49-74

59. Hartmann, P.E., and Prosser, C.G. (1984) Fed. Proc. 43, 2448-2453

60. Sharp, J.A., Lefevre, C., Watt, A., and Nicholas, K.R. (2016) Funct. Integr. Genomics 16, 297-321

61. Adel-Patient, K., Nutten, S., Bernard, H., et al. (2012) J. Agri. Food Chem. 60, 10858-10866

62. Qian, B., Xing, M., Cui, L., et al. (2011) J. Dairy Res. 78, 72-79

63. Dallas, D.C., Guerrero, A., Khaldi, N., et al. (2013) J. Proteome Res. 12, 2295-2304

64. Gifford, J., Hunter, H., and Vogel, H. (2005) Cell. Mol. Life Sci. 62, 2588-2598

65. Liepke, C., Zucht, H.-D., Forssmann, W.-G., and Standker, L. (2001) J. Chrom. B Biomed. Sci. App. 752, 369-377

66. Fiat, A.-M., Migliore-Samour, D., Jollès, P., et al. (1993) J. Dairy Sci. 76, 301-310

67. Chabancea, B., Jollèsa, P., Izquierdoa, C., et al. (1995) Br. J. Nutr. 73, 583-590

68. Teschemacher, H., Koch, G., and Brantl, V. (1997) Peptide Sci. 43, 99-117

69. Meisel, H., and FitzGerald, R.J. (2000) Br. J. Nutr. 84, 27-31

70. Migliore-Samour, D., Floc’h, F., and Jollès, P. (1989) J. Dairy Res. 56, 357-362

71. Gertrud, K., Klaus, W., and Hansjörg, T. (1985) Naunyn-Schmiedeberg’s Arch. Pharmacol. 331, 351-354

72. Wu, Z., Pan, D., Zhen, X., and Cao, J. (2013) J. Sci. Food Agri. 93, 1331-1337

73. Kanda, Y., Hisayasu, S., Abe, Y., Katsura, K., and Mashimo, K. (2007) Life Sci. 81, 449-457

74. Kumar, A., Buscara, L., Kuruppath, S., Ngo, K.P., Nicholas, K.R., and Lefèvre, C. In: Reyes Cruz, L.M., and Ortiz Gutierrez, D.C. (eds) Lactation: Natural Processes, Physiological Responses and Role in Maternity (2012) Nova Science Publishers, pp73-97

75. Modepalli, V., Kumar, A., Hinds, L.A., et al. (2014) BMC Genomics 15, 1012



Pesticides in Human Milk: Should We Be Concerned? Melvin Gay1*, Robert Trengove2,3 and Donna Geddes1

1School of Chemistry and Biochemistry, University of Western Australia, Crawley, WA 60092Separation Science and Metabolomics Laboratory, Murdoch University, Murdoch, WA 6150

3Metabolomics Australia, Murdoch University Node, Murdoch, WA 6150*Corresponding author: [email protected]

Breastfeeding has been the norm throughout primate evolution. Breastfeeding stimulates both the mother and the infant physiologically. Maternal effects include the release of oxytocin, which is essential for milk ejection but also induces a lower threshold to pain and stimulates positive feelings, hence its nick name the ‘love’ hormone. The skin-to-skin contact and the act of breastfeeding also modulates infant temperament, heart rate, temperature and acid–base balance. In addition, there are a myriad of positive effects on both the mother and infant that can in many cases be attributed to the unique species specific composition of human milk (HM) (1,2).

The short-term benefits of HM for the infant are extensive, where meta-analyses show a lower incidence of infection including necrotising enterocolitis (NEC), gastrointestinal tract infections and a reduction in infant mortality with increased doses of HM (3). Further, the lifetime programming effects of HM include a lower risk of obesity and diabetes later in life (4,5), which are precursors to metabolic disease as well as better neurological development and cognitive function compared to infants that were formula-fed (6,7). Indeed mothers also reap significant health benefits, particularly if they breastfeed their infants for more than 12 months, with a significant reduction in risk of cardiovascular disease, hypertension, diabetes and cancer (breast and ovarian) compared to those who have never breastfed (8-10).

The benefits of HM to infant are still therefore undeniable. However, the mechanisms associated with the transfer and/or production of nutritional and protective components in milk also act as pathways for the passage of environmental contaminants into the milk. Indeed persistent organic pollutants (POPs) and heavy metals have been detected in HM. The question is: does this pose a risk to the infant? Further, is the monitoring of these contaminants worldwide sufficient to safeguard not only the infants, but also everyone that lives in the same environment?

Persistent Organic PollutantsPOPs, such as pesticides, polychlorinated biphenyls

(PCBs) and polychlorinated dibenzo-p-dioxins (dioxins), are synthetic chemicals that are ubiquitously present in the environment (11,12). These pollutants are usually by-products from industrial processes or agrichemicals used for pest and weed control on farms. As many of these xenobiotics have lipophilic properties and are resistant to environmental degradation, they can be transported over a long distances via the atmosphere and water. Hence, these chemicals are then absorbed into mammals

via inhalation, ingestion and dermal contact, followed by bioaccumulation in organs with higher lipid contents, such as the liver and breast, as they are not easily metabolised in the body (12).

Pesticides, such as organochlorine pesticides (OCPs), organophosphate pesticides (OPPs) and carbamates, are used to control pests in agriculture and in homes. Besides their primary action as pesticides, many of these OCPs and OPPs can interfere with the functions of normal endocrine systems by mimicking, blocking, modulating or altering the synthesis, metabolism or transport of hormones (12,13). Long term pesticide exposure has also been associated with neuropsychiatric sequelae (14), chronic diseases such as Parkinson’s disease (15), cancer (16) and asthma (17), as well as congenital disorders (18,19).

While many of these pesticide groups, such as OCPs and OPPs, have been banned since the 1980s/90s due to their persistence and long half-lives (over seven years), many of these pesticides are still detected in the environment and in biofluids. A variety of pesticides, such as p,p’-dichlorodiphenyldichloroethylene (p,p’-DDE), p,p’-dichlorodiphenyltrichloroethane (p,p’-DDT) and hexachlorocyclohexane (HCH), have been detected in various biofluids of the inhabitants of different countries. p,p’-DDE, which is a metabolite of p,p’-DDT, is one of the most commonly detected OCPs in humans as DDT was widely used around the world in agriculture and domestic households in the 1940s/50s. Since the 1980s, many of these banned pesticides have been replaced by more ‘human-friendly’ pesticides, such as pyrethroids, which can be easily metabolised by mammals and have shorter half-lives of 2.5 to 12 hours in blood plasma (20).

As humans are not only exposed to a single chemical, but to a multitude of different chemicals simultaneously, it is important to continuously measure the level of environmental contaminants, both banned and legalised, to ensure that the total environmental pesticide exposure is within the safe threshold limits.

Impact of Pesticide Exposure on Infant HealthThe fetus and infants are recognised as being more

susceptible to the harmful effects of pesticides due to their small size and rapid growth as well as their immature immune and metabolic systems (21). Many of the studies designed to investigate the adverse effects of pesticides on infants have focused on maternal exposure to pesticides by measuring pesticide levels in maternal biofluids, such as blood (22,23), cord blood (24) and HM (25,26). Associations are then drawn between these measures and prenatal, infant and child development measures such as


Table 1. Concentrations of DDTs (ng/g fat) in human milk from various countries. Adapted from (25).

Country/ region

Asia

ChinaIndiaTaiwanKoreanJapanMalaysiaPhilippineTurkeyIran

Australian and New Zealand

New ZealandAustralia/WA

Europe

PolandSlovakiaNorwayLatviaDenmarkBelgiumGermanyCroatia/ZagrebUK

North and South Americas

USABrazil

Africa

South AfricaEthiopia

Year of sampling

2006/0720112000/0120082008/09200320042009

2006

2007/20102013/15

2002/0520032002/062002/041997/012009/102007/082009/102001/03

20042001/02

20012010

Mothers (number)

605336>509017334710

3940

28123771543845162054

3869

2839

Human milk samples

605336509#17334710

3740

28123771543845162954

3869

2829

Fat content (g/L)

313231223217223615

3933

142736nana44*36nana

22na

nana

DDTs (ng/g fat)

583a, *

1914b

333c

225c

119c

1600d

170b

338e

3560a

385f

63g

1621b

665d

53g

267h

82e

60*, d

125d

278b, *

157*, f

65a

493b

6320c

14460b

Reference

(52)(47)(53)(54)(55)(37)(56)(57)(36)

(58)(25)

(59)(60)(61)(62)(63)(64)(65)(66)(67)

(68)(69)

(70)(71)

na no data available* Values represented as median value# Pooled sample a Sum of o,p’-DDE + p,p’-DDE + p,p’-DDD + p,p’-DDTb Sum of p,p’-DDE + p,p’-DDD + p,p’-DDTc Sum of p,p’-DDE + p,p-DDD + o,p’-DDT + p,p’-DDTd Sum of p,p’-DDE + p,p’-DDTe Sum of o,p’-DDE + p,p’-DDE + o,p’-DDD + p,p’-DDD + o,p’-DDT + p,p’-DDTf Sum of o,p’-DDE + p,p’-DDE + p,p’-DDD + o,p’-DDT + p,p’-DDTg p,p’-DDE onlyh Sum p,p’-DDE + o,p’-DDT + p,p’-DDT

SHOWCASE ON RESEARCHPesticides in Human Milk



Pesticides in Human Milk

growth, immune and metabolic status. Studies have shown an association between higher

concentrations of the insecticide chlorpyrifos in maternal prenatal plasma and smaller infant birth weight and length (23). Similarly, smaller infant birth weight and head circumference have been observed in mothers with high prenatal urinary DDE concentrations (27). Further, recent studies have associated high exposure of OPPs with attention deficit/hyperactivity disorder in five-year-olds (28), and poor intellectual development and cognitive performance in seven-year-olds (29). Sex differences have also been identified where male infants were more susceptible to OPP exposure with smaller head circumference, delayed adaptive skills and social and motor development skills respectively (30). It is still difficult however to decide whether the associations are due to prenatal or postnatal pesticide exposure, therefore, one should exercise caution in the interpretation of these studies.

While many studies have used maternal pesticide levels and infant anthropometrical data at birth as a proxy measure of in utero development, our recent epidemiological longitudinal and cross-sectional studies have shown no significant associations between p,p’-DDE concentrations in milk from mothers in Western Australia and infant growth outcomes such as weight, length, head circumference and percentage fat mass (estimated using bioimpedance spectroscopy and ultrasound skinfolds) at 2, 5, 9 and 12 months postpartum (25). Furthermore, the concentration of p,p’-DDE significantly decreased by 68% from 70.9 ± 70.6 ng/g fat at 2 months to 22.4 ± 14.0 ng/g fat at 12 months (31), indicating that maternal bioburden is reduced via breastfeeding. Interestingly, expected relationships between maternal age (increased HM pesticide levels) and parity (reduced HM pesticide levels) are not always confirmed in the literature, however this is likely due to differences in study design and analysis. (22,25,27). One is tempted however, to speculate that the reduction of maternal bioburden during lactation could potentially contribute to the reported reduced incidence of breast and ovarian cancer in women that have breastfed (32-34). Whilst the Western Australia data is encouraging, the question is whether the dramatic reductions in POPs in HM are paralleled globally?

Temporal Trends and Worldwide Comparisons of Pesticides in HM

The ratio of p,p’-DDE to p,p’-DDT concentration indicates exposure history where a high DDE/DDT ratio (>5) represents historical exposure and a low DDE/DDT ratio (<5) suggests continuous exposure (35). This provides valuable insight into the maternal environment, lifestyle and dietary habits but also allows early intervention to minimiSe infant (and maternal) exposure to these pesticides if present at high concentrations. HM pesticide concentrations vary widely between countries (Table 1) and these are largely dependent on the extent of pesticide use and the timing of legal bans. Higher total DDT concentrations (sum of DDTs and its metabolites, DDE and DDD) are typically observed in malaria-prone countries such as Iran (36) and Malaysia (37), where DDTs were widely used to combat mosquitos and were only recently banned (1990s to 2000s). Limited use of DDT is also still allowed indoors in malaria endemic countries, such as South Africa and India (38). We have shown a declining trend in the total DDTs in HM from women in Western Australia since the 1970s (Fig. 1) (39-43). This is in agreement with Muller et al., where an annual decrease of approximately 12–13% of DDTs was observed in earlier years (1960s to 1980s) slowing to around a 5% annual decrease since the 1990s (40). On a national level, the HM p,p’-DDE concentrations observed in WA are 2.4 to 7.5 times lower than corresponding p,p’-DDE concentrations in other states and territories such as South Australia, Victoria and New South Wales, and is reflective of the inconsistent timing of legislation for pesticide control prior to the 1990s. Since levels of pesticides are declining in HM, it stands to reason infant dose is also declining.

Infant Daily Dose of Pesticides Via HMIt is essential to monitor HM pesticide concentrations to

ensure that the daily infant pesticide intake is within the tolerable limit of 0.5–10 µg/kg body weight/day (44-46) recommended by regulatory agencies (e.g. EPA, FAO, RIVM, WHO) to decrease the risk of impaired infant growth and development. Calculation of infant dose requires infant weight, concentration of the pesticide,

Fig. 2. Comparison between the actual daily intake (ADI) and estimated daily intake (EDI) of total DDTs by infants during the first year of lactation.

Fig. 1. Trends of total DDT concentrations in human milk of mothers in Western Australia.


milk fat and the volume of milk consumed. Many studies have used estimated values of milk fat, infant milk intake and infant weight, to estimate daily intake (EDI) of the infant (36,47,48). In our recent cross sectional study we calculated daily intake (CDI) by using the average milk intake based on the infant age (49) and measuring milk fat (25). This was further improved by calculating the actual daily intake (ADI) which is based on the actual measurements of milk fat, 24 hour milk intake and infant weight in our longitudinal study (Fig. 2) (31). The EDI significantly overestimates the overall daily intake over the first 12 months postpartum compared to both CDI and ADI by 53% and 44%. While Du et al. (31) observed a significant decrease in pesticide intake by the infant throughout the 12 months, they also observed that the ADI from several mothers exceeded the tolerable limit (>0.5 µg/kg body weight/day) in the earlier months of lactation (two and five months). This could be considered concerning, however this is counterbalanced by the fact that infant exposure to pesticides in HM via breastfeeding is a relatively short period as compared to a lifetime living in the same environment. Overall, the ADI we calculated was approximately three times lower than the tolerable limit. Thus, the benefits of HM, such as the immunological and nutritional benefits, far outweigh the potential negative effects that are yet to be adequately demonstrated. Therefore breastfeeding should continue to be encouraged for up to two years and beyond, as recommended by WHO (50).

Conclusions and Future DirectionsThe maternal environment, diet and body composition

before and during pregnancy play a major role in fetal programming and the future health of the infant (51). As part of a continuum, lactation represents a period of further infant programming for optimum health outcomes. Whilst the presence of pesticides in HM is of concern, the use of HM can provide a non-invasive avenue to monitor pesticide exposure and to measure infant daily intake. Further, while new data does not show detrimental effects of POPs in HM on early infant growth, longitudinal studies should be conducted for longer periods (beyond three years of life) to elucidate relationships with infant growth and developmental outcomes, including cognitive function. As many countries now source commodities, such as food produce, from various countries, it is essential that global restrictions are implemented and adhered to in order to reduce population contaminant exposure via diet.

References1. Newburg, D.S. (2001) Adv. Exp. Med. Biol. 501, 3-102. Hale, T.W., and Hartmann, P.E. (2007) Hale and

Hartmann’s Textbook of Human Lactation, 1st edition, Hale Publishing

3. Meinzen-Derr, J., Poindexter, B., Wrage, L., et al. (2009) J. Perinatol. 29, 57-62

4. Verduci, E., Banderali, G., Barberi, S., et al. (2014) Nutrients 6, 1711-1724

5. American Academy of Pediatrics. (2012) Pediatrics 129,

e827-8416. Newburg, D.S., and Walker, W.A. (2007) Pediatr. Res.

61, 2-87. Quigley, M. A., Hockley, C., Carson, C., et al. (2012) J.

Pediatr. 160, 25-328. Ip, S., Chung, M., Raman, G., et al. (2009) Breastfeed.

Med. 4 Suppl 1, S17-309. Schwarz, E.B., Ray, R.M., Stuebe, A.M., et al. (2009)

Obstet. Gynecol. 113, 974-98210. Stuebe, A.M., Willett, W.C., Xue, F., et al. (2009) Arch.

Intern. Med. 169, 1364-137111. Loganathan, B.G., and Kannan, K. (1994) Ambio 23,

187-19112. Tanabe, S. (2002) Mar. Pollut. Bull. 45, 69-7713. Bergman, A., Heindel, J.J., Kasten, T., et al. (2013)

Environ. Health Perspect. 121, A104-10614. Freire, C., and Koifman, S. (2013) Int. J. Hyg. Environ.

Health 216, 445-46015. Costa, L.G., Giordano, G., Guizzetti, M., et al. (2008)

Front. Biosci. 13, 1240-124916. Mostafalou, S., and Abdollahi, M. (2013) Toxicol. Appl.

Pharmacol. 268, 157-17717. Hernandez, A.F., Parron, T., and Alarcon, R. (2011)

Curr. Opin. Allergy Clin. Immunol. 11, 90-9618. Cavieres, M.F., Jaeger, J., and Porter, W. (2002)

Environ. Health Perspect. 110, 1081-108519. Weselak, M., Arbuckle, T.E., and Foster, W. (2007) J.

Toxicol. Environ. Health B Crit. Rev. 10, 41-8020. Leng, G., Leng, A., Kuhn, K.H., et al. (1997) Xenobiotica

27, 1273-128321. Bruckner, J.V. (2000) Regul. Toxicol. Pharmacol. 31, 280-

28522. Reid, A., Callan, A., Stasinska, A., et al. (2013) Sci. Total

Environ. 449, 208-21323. Perera, F.P., Rauh, V., Tsai, W.-Y., et al. (2003) Environ.

Health Perspect. 111, 201-20524. Ribas-Fito, N., Julvez, J., Torrent, M., et al. (2007) Am.

J. Epidemiol. 166, 1198-120225. Du, J., Gridneva, Z., Gay, M.C., et al. (2016)

Chemosphere 167, 247-25426. Ribas-Fito, N., Grimalt, J.O., Marco, E., et al. (2005)

Environ. Res. 98, 8-1327. Wolff, M.S., Engel, S., Berkowitz, G., et al. (2007)

Pediatr. Res. 61, 243-25028. Marks, A.R., Harley, K., Bradman, A., et al. (2010)

Environ. Health Perspect. 118, 1768-177429. Bouchard, M.F., Chevrier, J., Harley, K.G., et al. (2011)

Environ. Health Perspect. 119, 1189-119530. Liu, P., Wu, C., Chang, X., et al. (2016) Environ. Health

Perspect. 124, 1637-164331. Du, J., Gridneva, Z., Gay, M. C., et al. (2016) Sci. Rep. 6,

3835532. Danforth, K.N., Tworoger, S.S., Hecht, J.L., et al. (2007)

Cancer Causes Control 18, 517-52333. Collaborative Group on Hormonal Factors in Breast

Cancer (2002) Lancet 360, 187-19534. Shema, L., Ore, L., Ben-Shachar, M., et al. (2007) J.

Cancer Res. Clin. Oncol. 133, 539-54635. Mishra, K., and Sharma, R.C. (2011) Sci. Total Environ.

409, 4939-4949

SHOWCASE ON RESEARCHPesticides in Human Milk


36. Behrooz, R.D., Sari, A.E., Bahramifar, N., et al. (2009) Chemosphere 74, 931-937

37. Sudaryanto, A., Kunisue, T., Tanabe, S., et al. (2005) Arch. Environ. Contam. Toxicol. 49, 429-437

38. Mandavilli, A. (2006) Nat. Med. 12, 870-87139. Noren, K., and Meironyte, D. (2000) Chemosphere 40,

1111-112340. Mueller, J.F., Harden, F., Toms, L.-M., et al. (2008)

Chemosphere 70, 712-72041. Stacey, C., Stanley, W., and Whitney, S. (1985) Arch.

Environ. Health 40, 102-10842. Stacey, C.I., and Thomas, B.W. (1975) Pestic. Monit. J.

9, 64-6643. Stevens, M.F., Ebell, G.F., and Psaila-Savona, P. (1993)

Med. J. Aust. 158, 238-24144. FAO/WHO (2000) 45. RIVM (2001) Re-evaluation of human-toxicological

maximum permissible risk levels. National Institute of Public Health and the Environment, Bilthoven, The Netherlands

46. USEPA (1996) Integrated Risk Information System (IRIS). DDT: non-cancer assessment

47. Bedi, J.S., Gill, J.P.S., Aulakh, R.S., et al. (2013) Sci. Total Environ. 463-464, 720-726

48. Van Oostdam, J., Gilman, A., Dewailly, E., et al. (1999) Sci. Total Environ. 230, 1-82

49. Kent, J.C., Mitoulas, L., Cox, D.B., et al. (1999) Exp. Physiol. 84, 435-447

50. World Health Organization, and UNICEF. (2003) Global strategy for infant and young child feeding, WHO, Geneva, Switzerland

51. Godfrey, K.M., and Barker, D.J. (2001) Public Health Nutr. 4, 611-624

52. Leng, J.-H., Kayama, F., Wang, P.-Y., et al. (2009) Chemosphere 75, 634-639

53. Chao, H.-R., Wang, S.-L., Lin, T.-C., et al. (2006) Chemosphere 62, 1774-1785

54. Kim, D., Ryu, H.Y., Lee, J.H., et al. (2013) J. Environ. Sci. Health B 48, 243-250

55. Fujii, Y., Haraguchi, K., Harada, K.H., et al. (2011) Chemosphere 82, 25-31

56. Malarvannan, G., Kunisue, T., Isobe, T., et al. (2009) Environ. Pollut. 157, 1924-1932

57. Çok, I., Mazmanci, B., Mazmanci, M.A., et al. (2012) Environ. Int 40, 63-69

58. Mannetje, A., Coakley, J., Bridgen, P., et al. (2013) Sci. Total Environ. 458–460, 399-407

59. Hernik, A., Góralczyk, K., Struciński, P., et al. (2011) Ann. Agric. Environ. Med. 18, 113-118

60. Yu, Z., Palkovicova, L., Drobna, B., et al. (2007) Chemosphere 66, 1012-1018

61. Polder, A., Skaare, J.U., Skjerve, E., et al. (2009) Sci. Total Environ. 407, 4584-4590

62. Bake, M.A., Linnika, Z., Sudmalis, P., et al. (2007) Int. J. Hyg. Environ. Health 210, 483-489

63. Shen, H., Main, K.M., Virtanen, H.E., et al. (2007) Chemosphere 67, S256-S262

64. Croes, K., Colles, A., Koppen, G., et al. (2012) Chemosphere 89, 988-994

65. Raab, U., Albrecht, M., Preiss, U., et al. (2013) Chemosphere 93, 461-467

66. Klinčić, D., Romanić, S.H., Sarić, M.M., et al. (2014) Environ. Toxicol. Pharmacol. 37, 543-552

67. Kalantzi, O.I., Martin, F.L., Thomas, G.O., et al. (2004) Environ. Health Perspect. 112, 1085-1091

68. Johnson-Restrepo, B., Addink, R., Wong, C., et al. (2007) J. Environ. Monit. 9, 1205-1212

69. Azeredo, A., Torres, J.P.M., de Freitas Fonseca, M., et al. (2008) Chemosphere 73, S246-S251

70. Darnerud, P. O., Aune, M., Larsson, L., et al. (2011) Sci. Total Environ. 409, 4048-4053

71. Gebremichael, S., Birhanu, T., and Tessema, D.A. (2013) Chemosphere 90, 1652-1657


Pesticides in Human Milk


Medallist and Awardee ProfilesThe Lemberg Medal is awarded to a distinguished Australian biochemist or molecular biologist who will present the Lemberg Lecture at the ComBio meeting. The Medal is presented in memory of Emeritus Professor M.R. Lemberg, who was the Society’s first President and Honorary Member. Nominees must have been members of the Society for at least five years before the year in which the Medal nomination is to be considered. An honorarium is provided by ASBMB.

The Lemberg Medal:

John Mattick

John Mattick is the Executive Director of the Garvan Institute of Medical Research. He obtained his BSc with First Class Honours in Biochemistry from the University of Sydney, and his PhD in Biochemistry from Monash University, under the supervision of Professor Tony Linnane and Professor Phillip Nagley. He then undertook postdoctoral training with Professor Salih Wakil at Baylor College of Medicine in Houston, where his work on the architecture of the fatty acid synthase complex is now featured in biochemistry textbooks.

John returned to Australia in 1982 to join the (then) CSIRO Division of Molecular Biology, where he developed one of the world’s first genetically engineered vaccines (against ovine footrot).

In 1988, he joined the University of Queensland as the Foundation Professor of Molecular Biology and Director of the Centre for Molecular Biology and Biotechnology, which later became the ARC Special Research Centre for Molecular and Cellular Biology, and ultimately the Institute for Molecular Bioscience (IMB). John was also the Foundation Director of the Australian Genome Research Facility (1996–2002) and the ARC Special Research Centre for Functional and Applied Genomics.

In 2006, John relinquished the IMB Directorship to take up an ARC Federation Fellowship (and later an NHMRC Australia Fellowship) to work on his longstanding conviction that the vast amounts of untranslated RNAs that are expressed from the genomes of complex organisms are not junk, but rather an extensive regulatory system responsible for guiding the epigenetic trajectories of development.

He had first published his hypothesis after a sabbatical in Cambridge in 1994, and in the ensuing two decades, showed the extensive transcription of intronic, intergenic and antisense RNAs, their highly dynamic and specific cellular and subcellular expression, their association with chromatin-modifying complexes and their involvement in development and disease. He also discovered

ultraconserved elements in the human genome and several classes of small RNAs, and, in conjunction with colleagues, developed new methods for high-resolution transcriptome analysis.

John returned to Sydney in 2012, where he established the Garvan Institute as one of the world’s leading centres for human genome sequencing and oversaw the launch of Genome.One, a wholly owned subsidiary of the Garvan and one of the world’s first clinically accredited whole genome sequencing companies.

John has published over 300 papers, which have been cited over 32,000 times (Scopus). Inter alia, he has been Chair of the Queensland Studies Authority and a member of the Australian Health Ethics Committee. John was recently named by the NHMRC as the one of the all-time high achievers in Australian health and medical research.

The Merck Research Medal is awarded to an outstanding Australian biochemist or molecular biologist with less than 15 years postdoctoral experience. The successful candidate will present the Merck Medal Lecture at the ComBio meeting. Nominees must have been members of the Society for at least two years before the year in which the Medal nomination is to be considered. An honorarium is provided through the generosity of Merck Pty Ltd.

The Merck Research Medal:

Lars Ittner

Professor Lars Ittner is a group leader in the Department of Anatomy of the Faculty of Medicine at the University of New South Wales (UNSW) and Senior Principal Research Fellow at Neuroscience Research Australia (NeuRA) in Sydney. He is a neuroscientist and heads the research laboratories of the Dementia Research Unit and the Transgenic Animal Unit of the Mark Wainright Analytical Centre at UNSW.

Lars received his medical degree from the University of Ulm in Germany in 2001, followed by an experimental thesis in medical research at the University of Zurich in Switzerland in 2002. Lars was then selected into the Postgraduate Course for Experimental Biology and Medicine at the University of Zurich, Switzerland, from where he continued as a postdoctoral fellow with Professor Jan A. Fischer, working on G-protein-coupled receptors and neuronal development.

In 2005, Lars moved to Australia to work as a Postdoctoral Fellow with Professor Jürgen Götz at the University of Sydney. With this move, Lars changed his research focus from early neuronal development to neurodegenerative diseases, which continues to be the major interest of his research program. During his time at the University of


Medallist and Awardee ProfilesSydney, he made fundamental discoveries about the synaptic functions of one of the hallmark proteins in Alzheimer’s disease, the tau protein. Furthermore, he developed a range of novel transgenic mouse models of neurodegenerative diseases that continue to be used for understanding pathomechanisms and testing of novel treatments. Lars became a Senior Lecturer and Associate Professor at the University of Sydney’s Brain and Mind Research Institute in 2009 and 2011, respectively.

Lars moved to UNSW in 2013, where he was appointed as Professor in the School of Medical Sciences to head the Dementia Research Unit. His current work aims to understand the earliest changes at the synapse of neurons during the onset of Alzheimer’s disease. Furthermore, Lars works together with industry partners to translate his findings into therapeutic avenues of the future. Lars is also part of a large chief investigator team, dedicated to understanding and treating rarer forms of dementia and motor neuron disease.

Lars has published over 100 peer-reviewed articles in journals, including Science, Cell, Nature Medicine, Cell Stem Cell, Nature Reviews Neuroscience, Acta Neuropathologica and PNAS. He has held prestigious fellowships throughout his career, including a NHMRC Senior Research Fellowship, and he has been awarded multiple Project and Program Grants from both the NHMRC and ARC as chief investigator. Amongst several awards he has received for his research, he has also been awarded the Merck Young Achiever Award from the Australian Society for Medical Research (ASMR) in 2010.

The Shimadzu Education Award rewards outstanding achievement in education of biochemistry or molecular biology, especially innovation and creativity in education, with a view to fostering leadership in this important area of the Society’s objectives. The Award will enable the recipient to participate in an international conference with a significant focus on education, or to spend a period of time at another institution (in Australia or overseas) for the purposes of undertaking developments in education of biochemistry and molecular biology. The recipient will present a lecture within the Education Symposium at the ComBio meeting. Applicants must have been members of the Society for at least two years before the year in which the Award application is to be considered. The contribution to travel expenses is provided through the generosity of Shimadzu Excellence in Science Program.

The Shimadzu Education Award:Heather Verkade

Heather Verkade’s interest in science began at the University of Melbourne, with a Bachelor of Science and

honours in genetics. She carried out a PhD at the Peter MacCallum Cancer Institute (back in East Melbourne) in yeast cell cycle genetics, but she then changed direction to study zebrafish developmental genetics at UCSF in San Francisco and then at the Ludwig Institute. She has always loved to study complex systems, and to consider both the detail of a reductionist approach at the same time as the effect on the whole organism. Heather gained a teaching and research academic position at Monash University in the School of Biological Sciences, but realised that she was most fascinated with university education and the complex processes by which students learn. In 2014, she was fortunate to gain a Teaching Specialist position as a Senior Lecturer at the University of Melbourne in the Department of Biochemistry and Molecular Biology.

Heather believes that student-centred activities are the key to quality teaching. She aims to teach in a way that results in students being actively engaged in thinking about (and discussing!) molecular biology problems in the classroom, even though those classrooms are sometimes very large. This approach is designed to make students apply their understanding, even if at a basic level and without a laboratory, to enable students to fully internalise knowledge. Students in Heather’s classes are often trying to interpret experimental data or design simple experiments. She trusts that students are ready to try challenging activities with higher order thinking, as long as they are trained and supported and the expectations are clear and consistent.

Heather has been researching ways to confront student misconceptions in the classroom in a way that allows them to rethink their understanding. She has been leading a research project funded by a University of Melbourne Learning and Teaching Initiative grant, to trial different ways to change student misconceptions in a range of different science, technology, engineering and mathematics (STEM) fields, and a range of different educational situations. She also runs an Education Research Group within the School of Biomedical Sciences, to support academics who wish to carry out Scholarship of Teaching and Learning (SoTL) and be recognised for their achievements and innovations. One aim of this is to promote the roles of Teaching Specialist academics within the university.


The Beckman Coulter Discovery Science Award is awarded to an Australian biochemist or molecular biologist for distinguished contributions to the field of biochemistry and molecular biology. The Award is intended as a Travelling Lectureship to enable the awardee to present his/her work at a number of centres within Australia and New Zealand. Nominees must have been members of the Society for at least two years before the year in which the Award nomination is to be considered. The award carries an honorarium to cover the travelling expenses, provided through the generosity of Beckman Coulter.

The Beckman Coulter Discovery Science Award:Mike Lawrence

I began my research career in 1978, undertaking a PhD degree in theoretical physics at the University of Cape Town (UCT) in South Africa. My thesis concerned the semi-classical modelling of the quantum mechanics of protein tunnelling in hydrogen-bonded compounds, and it was heavily based on my mathematics and physics background. However, I was always fascinated by the application of these disciplines to the life sciences and in 1980, I was fortunate to secure funds for a two-year postdoc at the MRC Laboratory for Molecular Biology in Cambridge, UK. This was a turning point in my career and I found myself on a very steep learning curve! During the postdoc I determined the structure of the T-state of the allosteric enzyme phosphofructokinase from Bacillus stearothermophilus and developed techniques in electron micrograph image alignment, some of which are still used today in cryo-tomography.

I then returned to South Africa for five years. Unfortunately, South Africa had no resources at the time for structural biology, but nevertheless I secured the directorship of a small electron microscopy facility within the science faculty at UCT and had quite a bit of fun.

In 1988, I emigrated to Australia and joined the research group headed by Peter Colman, which proved to be a further turning point of my career. During my eighteen years at CSIRO, I worked on a number of particularly challenging projects in structural biology, including determination of the structure of phaseolin (a seed storage protein), the fusion protein of Newcastle disease virus (NDV-F) and, ultimately, the extracellular domain of the human insulin receptor, in a project led by Dr Colin Ward. The insulin receptor project terminated in 2006 and I decided to accept a position as a laboratory head at the Walter and Eliza Hall Institute of Medical Research (WEHI) in Parkville, Victoria, to continue the quest to determine how insulin bound to and activated

its receptor. This opportunity was undoubtedly the most exciting in my career, not only because it offered me significant independence but also because it gave me the opportunity to pursue a very important and still unsolved problem in structural biology, with direct application to human health. The WEHI has proved to be an incredibly supportive and stimulating environment in which to work. In 2013, my lab finally obtained the first atomic picture of insulin bound to its receptor and published it in Nature. Since then my laboratory has sought to build on this finding, not only to fill in the many gaps remaining but to pursue active ways of translating it into better insulins and into applications in cancer.

The Eppendorf Edman Award is awarded to a biochemist or molecular biologist with no more than seven years postdoctoral experience, in recognition of their outstanding research work. The Award provides funds to assist the recipient to attend an overseas conference in a field associated with biochemistry or molecular biology or to briefly visit a research laboratory in Australia or elsewhere to access specialised equipment or to learn new research techniques. Applicants must have been members of the Society for at least 12 months before the year in which the Award application is to be considered. The contribution to travel expenses is provided through the generosity of Eppendorf South Pacific.

The Eppendorf Edman Award:

Natalie Spillman

During my research-focussed Bachelor of Philosophy degree at the Australian National University, I developed an interest in parasitology, and have spent my career studying the malaria parasite. Living inside a host red blood cell presents many challenges to the parasite, and the tricks and unique biology it uses to thrive in this environment keep me fascinated every day. I completed my PhD at ANU in 2012 under the supervision of Professor Kiaran Kirk and Dr Richard Allen, investigating Na+ and pH regulation in the malaria parasite. Although I was interested in Na+-transport as an elementary and crucial aspect of cellular physiology, I demonstrated that a key transporter involved in parasite Na+-regulation, a P-type Na+-ATPase called PfATP4, was involved in resistance to a suite of new antimalarial drugs. My PhD work sparked a flurry of work in the malaria field concerning PfATP4 and new compounds targeting PfATP4.

Following this, for my postdoctoral work I switched from cell biology techniques studying ion fluxes, to using biochemical analyses and genetic manipulation to understand how the parasite modifies and alters its red blood cell host. I also switched continents, and moved

Medallist and Awardee Profiles


to Washington University School of Medicine, in Saint Louis, Missouri, USA. Under the mentorship of Professor Daniel Goldberg I investigated how the malaria parasite alters erythrocyte signalling lipids, and hypothesised why this is important for disease pathogenesis. My overseas postdoctoral studies were supported by two competitive fellowships, the Australia to USA Fellowship from the American Australian Association and an NHMRC CJ Martin Overseas Biomedical Early Career Fellowship.

In 2016, I returned to Australia and moved to the Biochemistry and Molecular Biology department at the University of Melbourne. Here I am part of the laboratory of Professor Leann Tilley, working on untangling the molecular mechanisms of drug action and drug resistance. In the future, I hope to combine my interests in cell physiology, membrane signalling and drug resistance to continue to unravel the unique biology of the malaria parasite.

I am honoured and excited to be the recipient of the 2017 Eppendorf Edman Award, which will allow me to present my current research at the EMBO/EMBL Molecular and Cell Biology of Membranes meeting in Heidelberg, Germany.

The Boomerang Award is offered to an outstanding expatriate Australian biochemist or molecular biologist to provide an opportunity to return to Australia to present their work in a symposium at the ComBio meeting and to give seminars at universities or research institutes. This will provide the awardee with exposure in Australia and will facilitate interactions with local researchers. The Award will include free registration at ComBio, as well as significant contributions to travel and accommodation expenses. Applicants must have been a member of a recognised Australian scientific society for at least two years, and be no more than seven years since the award of their PhD. The contribution to travel expenses is provided by ASBMB.

The Boomerang Award:

Yan Yan Yip

Yan Yip’s first exposure to biomedical research was at the Department of Pathology, the University of Melbourne, as an Honours student under the supervision of Dr Wendy Cook. She studied the post-transcriptional regulation of a novel gene that is silenced in acute myeloid leukaemia, learning a variety of molecular biology techniques along the way.

Yan then joined the laboratory of Dr Amardeep Dhillon at the Department of Biochemistry and Molecular Biology located within the Bio21 Molecular Science and Biotechnology Institute as a research assistant. Here, she helped identify key determinants of Raf-1 kinase activity,

a component of the Raf/MEK/ERK signalling cascade that is often deregulated in cancer. This experience not only expanded her repertoire of cell biology and biochemistry techniques but also kick-started her interest in cell signalling.

In 2010, Yan started her PhD under the guidance of Dr Dominic Ng and Associate Professor Marie Bogoyevitch, where she aimed to characterise signalling pathways that target the microtubule network, specifically in the context of cellular stress. She discovered that stress stimulation activates both the c-Jun N-terminal kinase and cyclic-AMP protein kinase pathways, which then act synergistically to protect the microtubule network from depolymerisation by phosphorylation and subsequent inactivation of stathmin, a microtubule destabilising protein. This work, published in the Journal of Biological Chemistry, highlights a mechanism by which the cell is able to adapt to deleterious stress stimuli in order to maintain cell viability.

Keen to continue working on aspects of microtubule function, Yan joined the laboratory of Dr Mark Dodding at the Randall Division of Cell and Molecular Biophysics, King’s College London for her postdoctoral work. The key research interest of the Dodding lab is to understand the molecular basis of microtubule-based intracellular cargo transport, having solved the first crystal structure of the interface between the kinesin-1 microtubule motor protein and its cargo. In collaboration with Dr Roberto Steiner, also at the Randall, Yan found that the light chains of kinesin-1 undergo a cargo binding-induced conformational change that confers activity to the holoenzyme. Identification of this novel molecular switch extends our understanding of how cargo selectivity can impact kinesin-1 function and this work was recently published in PNAS.

Yan is grateful to the ASBMB for the Boomerang Award. She will use this opportunity to present her work at ComBio2017 in Adelaide and to engage with research labs in Brisbane and Melbourne.

Medallist and Awardee Profiles


SAMUEL ROBINSON - recipient of the Fred Collins Award for the most outstanding ASBMB Fellowship applicant

Samuel Robinson completed his BSc and BSc (Honours) at the University of Auckland, New Zealand. In 2011, he received an Australian Postgraduate Award, to undertake a PhD at Monash University, supervised by Professor Ray Norton. Since the completion of his PhD in 2015, for which he was awarded the Mollie Holman medal, he has been based at the University of Utah, USA, working as a postdoctoral fellow in the lab of Professor Baldomero Olivera. This year he will return to Australia to join Professor Glenn King and Dr Eivind Undheim at the University of Queensland.

His research interest is in understanding the composition and mechanism of action of animal venoms. A major research highlight has been his pioneering of the use of proteogenomics for the identification of new, previously hidden, classes of bioactive

peptides. One outcome of this was the discovery that a class of specialised insulins form an integral part of the venoms of some species of marine cone snail. This provided ground-breaking insight into the structure-activity relationship of insulin, and has led to the preclinical development of a new anti-diabetes agent. This research was featured by numerous newspapers and science magazines around the world, including The Washington Post, The Guardian, New Scientist, Scientific American and National Geographic.

This ASBMB Fellowship will allow him to spend time working in the lab of Dr Beatrix Ueberheide, a world expert in mass-spectrometry, at New York University, USA, where he will aim to expand his knowledge and skillset in the identification and characterisation of venom peptides.

ASBMB FELLOWSHIP PROFILES

The ASBMB Fellowships are awarded annually to biochemists or molecular biologists, in their early career and normally resident in Australia, in recognition of their outstanding work in an area of biochemistry and molecular biology.The Fellowships provide funds to assist the recipient to attend an overseas

conference in a field associated with biochemistry or molecular biology or to briefly visit a research laboratory in Australia or elsewhere to access specialised equipment or to learn new research techniques.Applicants must be at least in the second year of PhD training and not more than

2 years subsequent to the award of the PhD degree. Applicants must have been members of the Society for least 1 year immediately prior to application.

GEORGIA ATKIN-SMITHGeorgia Atkin-Smith completed a Bachelor of Biotechnology and Cell Biology at La

Trobe University in 2013. She then completed her honours in Biochemistry under the supervision of Dr Ivan Poon and Dr Mark Hulett, finishing with first class honours. During this year, Georgia began to work on the molecular mechanisms and functions of apoptotic cell disassembly in monocytes, demonstrating a novel mechanism of disassembly which resulted in a first author Nature Communications paper.

Now in the third year of her PhD, Georgia has continued her work on the disassembly of apoptotic monocytes with a first author Scientific Reports manuscript characterising a novel method to isolate apoptotic bodies. Georgia has also published a review in Trends in Cell Biology and contributed to publications in Nature Protocols and Apoptosis. Current unpublished findings include the identification of the first positive regulator of apoptotic cell disassembly and the elucidation of the role of disassembly in infection in both viral

propagation and immune detection. Recently, Georgia was a finalist for the Research Impact Victorian Young Achievers Award 2016 and was awarded

the overall People’s Choice. Additionally, after a two week research placement in a laboratory in Tokyo, Japan, Georgia was invited to present her current research findings at four universities throughout Tokyo and has also been selected for numerous domestic conference oral presentations.

With the help of this ASBMB Fellowship, Georgia is planning to travel to Toronto, Canada, this year to present her research at the International Society of Extracellular Vesicles Annual Conference.


Publishing your First first-author paper

You have worked hard in the lab and your supervisor thinks you have a nice story to tell. Job done, right? No, now it is time to convince your academic peers that your work is worth publishing in a peer-reviewed journal. Your research is only going to reach others if it is being communicated effectively and publishing in a journal is a good starting point. To be a successful researcher, not only do you need to have motivation, perseverance, time management and technical skills but you also need to be able to write well. Writing a scientific paper is something that comes naturally to some, but not to others. It may seem like a daunting task and a good paper may take a very long time to write, but hopefully the following tips can help you with the process (which I am still learning myself!).

ASBMB FELLOWSHIP PROFILES

YILIN KANGYilin Kang obtained a Bachelor of Science with First Class Honours in 2014 from the

University of Melbourne, majoring in Biochemistry and Molecular Biology. In the final year of her undergraduate degree, Yilin was exposed to mitochondrial protein biogenesis in the lectures of Dr Diana Stojanovski, a mitochondrial biologist from the Bio21 Institute. Mitochondria immediately caught Yilin’s interest and she was fortunate enough to undertake a mini project in the lab of Dr Stojanovski, where she developed a passion for mitochondrial research. Yilin stayed on with Diana for her Honours project in 2014 and is now in the third year of her PhD.

Her doctoral studies have focused on the characterisation of a human mitochondrial protein import machinery – the Translocase of the Inner Membrane 22 (TIM22) complex. Although protein transport systems are well-studied in yeast, the human TIM22 complex were still poorly characterised when Yilin commenced her PhD. Yilin’s work has uncovered

a novel, metazoan-specific subunit of the human TIM22 complex, Tim29, which has a role in mediating the assembly and stability of the mature TIM22 complex. Identification of Tim29 highlights the importance of analysing protein import systems across phylogenetic boundaries, which can reveal novel facilitators and mechanisms in protein trafficking. Yilin’s work on Tim29 was published in Elife in 2016 and she has presented her research at several conferences including AussieMit 2016, where she received a Young Investigator Award.

This ASBMB Fellowship will allow Yilin to attend and present her studies on mitochondrial protein translocation at the 2017 EMBO Conference on Protein Translocation and Cellular Homeostasis in Dubrovnik, Croatia.

STEPHANIE BEGGStephanie completed a Bachelor of Science (Biomedical Science) at the University of

Adelaide in 2012, majoring in Microbiology, Immunology, and Biochemistry. Following her undergraduate studies, she was accepted into the accelerated Masters of Philosophy program within the Research Centre for Infectious Diseases (RCID), under the supervision of Associate Professor Christopher McDevitt and Professor James Paton. She was later upgraded to a full PhD to further her research into the human bacterial pathogen Streptococcus pneumoniae. Her thesis investigated this bacterium’s cellular requirement for essential trace metals, such as manganese and zinc, and focussed on how S. pneumoniae steals these metals directly from the host environment. This scavenging by invading bacteria is crucial for both their survival and ability to cause disease within humans. Her research delivered both first author and co-authored publications in journals such as Nature Chemical Biology, Nature Communications and ACS Chemical Biology. Stephanie was awarded

the University Doctoral Research Medal upon completion of her PhD in 2016, and is now a postdoctoral researcher in the RCID. She is currently investigating the molecular details of how metal poisoning and starvation impact S. pneumoniae, as these processes are directly relevant to how the human immune system attempts to kill invading bacteria. This ASBMB Fellowship will allow Stephanie to attend the Cell Biology of Metals Gordon Research Conference in Vermont, USA, in July 2017, and present her recent research into the prokaryotic metalloproteome to an international audience.

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415-Australian.Biochemist.10.10.16

1. Decide on your main messageWriting a paper is merely a way to tell people what you have discovered.

Decide on what story you want to tell and what the main point of your paper will be. Then you pick the results that best help you tell your story. List all your results and decide on whether you have enough ‘evidence’ to draw your conclusions. It can be a bit disheartening when you realise that only a small fraction of your results actually make it into a paper. You should resist the temptation to try and cram in as many results as possible in order to tell multiple stories in one paper. The key is to put yourself in the reader’s shoes to get your message across.

2. Pick a journalOnce you have decided on your take home message, decide on which

journal you want to submit your article to. Your supervisor and collaborators will be able to help you decide on the most appropriate journal. Finding the ‘right’ journal can be difficult and you may want to take into account the research area, reputation and target audience of the journal. Assessing the quality of a journal by its impact factor is a controversial issue as there are other factors that can influence the impact factor, not necessarily related to the quality of the papers being published. Nevertheless, the impact factor is still one of the main criteria to consider when choosing a journal. It is always a good idea to check which journals publish work similar to yours. You typically aim high (more prestigious journals) to begin with because you can resubmit to less prestigious journals if you get rejected. Be honest with yourself about where you realistically can see your work getting published, otherwise you may lose valuable time (and sleep) reformatting papers for submission. Chances are that your first paper will not be published in Nature or Science – many might go through their whole careers without ever getting one! So don’t be disheartened as plenty of influential, highly cited and high-quality papers are published outside of these journals. Consider open access policies for the journals and publication fees. Once you have picked the journal, read the guidelines for authors that you must adhere to.

3. Prepare your figuresWhenever I get some exciting new data, I try to generate publication-

quality figures as soon as possible, which can then be used in presentations and manuscripts. A well-constructed figure is worth a thousand words. Of course your figures do need to abide by the target journal’s guidelines, so make sure you can still edit them if need be. The figures and tables, including the legends, should be self-explanatory and the information conveyed in a figure should not be repeated in another figure. I also find it is a good idea to have a mixture of different types of figures for a paper. Any figures that don’t add to the main story but are important in supporting the paper can be included in the supplementary material. Don’t forget to include appropriate axes titles and units.

4. Write a cohesive story – introduction, methods, results and discussionIt is unlikely that you will be writing your first paper alone. Remember

that the reviewers are not necessarily in your niche area of research and you should make the review experience as painless as possible for them. I find it useful to find a good paper in the same journal that you can use as a guide.

A good introduction should convince readers that your work is important and will address an unresolved problem or gap in the literature. Add enough background information so that the reader can understand the remainder of the paper.

The methods you used in your paper should include enough detail that a fellow researcher can reproduce the experiment. But if a method is well established, use references to previously published procedures to keep this section succinct.


Once you have prepared your figures and tables, you will find the results section relatively straightforward to compile. I tend to group the results together into subheadings if this is permitted by the journal. Keep the data in a logical order that helps you tell a coherent story, which is not necessarily the order you performed your experiments in.

After you have presented your results, it is in the discussion that you highlight the importance of your data and explain what it all means. You need to put your data into the context of your field. When writing your discussion, try not to overstate the conclusions. Try to come up with all plausible explanations for your results and let the results speak for themselves.

The order in which you write these sections is up to you. One of the key points in writing a good paper is ensuring that one section flows into the next.

5. Title and abstractEven though a title is only a few words long, it often takes time to come up

with one that you are happy with. Keep the title succinct and descriptive of the content but not so technical that it cannot be easily understood. I know a number of people who make the decision to read a paper or not based on the title. Think of the title as the advertisement of your article.

The abstract should be succinctly written and avoid the use of jargon. It is a way of highlighting the aim of the paper and the main findings. In some journals, the editor may only look at the abstract to decide whether a paper should be further considered and sent for review.

6. Feedback

Try not to dwell on your first draft too much before sending it to your supervisor for revision. More often than not, the draft will come back to you covered in red writing. Once you have incorporated your supervisor’s feedback, pass it on to your co-authors. It is a requirement that all authors on the papers have revised and approved the final version of the manuscript. I also find it useful to get someone outside of my discipline or even a non-scientist to read the final draft of the paper. It is unbelievable the errors they can pick up even when they only understand the meaning of a few words.

7. Submission and revisionThe senior author on the paper will be in charge of submitting the revised

draft of the paper alongside a cover letter. The review process may take a few days but more likely, a few months. If major/minor changes are required you will have a timeframe to resubmit the paper. Consider the reviewers’ comments carefully as you will often have to prepare a letter to respond to their feedback. When preparing your response letter, make sure you carefully address each comment in a calm and methodical way. I always find it a good idea not to address the comments immediately after reading it for the first time. Also, don’t think of the comments as a personal attack on your research but merely a way of improving the quality of your publication. A glass of wine may also ease the process! Sometimes, reviewers may request changes to the manuscript or even more data. If you decide not to follow the request, make sure you provide a good justification for not doing so. Simply ignoring a request or criticism will not be looked upon favourably by the editor. Once you have addressed the reviewers’ comments, with any luck, your paper will be accepted by the journal. When your work gets published in a journal, you tend to forget about the pain you had to endure during peer review and you start looking forward to putting together your next publication!

The Student’s Page is coordinated by Dr Tatiana Soares da Costa, who is an NHMRC Early Career Fellow at the La Trobe Institute for Molecular Science ([email protected]).

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Australia Day Honours for ASBMB MembersProfessor David Vaux

was awarded an Officer of the Order of Australia (AO) for ‘distinguished service to medicine in the field of biomedical cancer research, to higher education as an academic and mentor, and to professional integrity and ethics.’

Dave Vaux (or ‘Davo’) is currently Deputy Director of the Walter and Eliza Hall Institute (WEHI). He trained in medicine at the University of Melbourne,

spending a year in the middle of his degree to do research in Gus Nossal’s lab. He returned to WEHI after his intern year at the Royal Melbourne Hospital to undertake a PhD studying B cell malignancies supervised by Jerry Adams.

Working on the lymphoma associated Bcl2 gene, he showed that unlike other cancer genes known at the time, it did not promote cell growth and proliferation, but stopped cells from being able to kill themselves. This showed that cell division and cell death were separately regulated independent processes. Bcl2 thus became the first component of the cell death mechanism to be recognised. Because of the association of Bcl2 translocations with lymphoma, it also became clear that failure of cell death could promote malignancies in humans, and avoidance of apoptosis is now recognised as one of the ‘hallmarks of cancer’.

He showed that Bcl2 could synergise with the c-myc oncogene to transform cells in vitro, and by making Bcl-2 transgenic mice and crossing them with myc transgenic mice, he showed they could also synergise to cause leukemia in vivo.

In 1989 he went to Stanford, and together with Stuart Kim made transgenic C. elegans worms expressing Bcl2, and showed that the mechanism for apoptosis of mammalian cells and the mechanism for programmed cell death in the worm were evolutionarily conserved. Expression of human Bcl2 in the worms prevented most of the cell deaths that were normally programmed to occur during development, and showed that the mechanism for apoptosis of mammalian cells and the mechanism for programmed cell death in the worm were evolutionarily conserved.

His association with biotech companies IDUN and TetraLogic helped in the translation of discoveries of the basic biology of cell death into drugs that inhibit the Bcl2 and IAP families of proteins.

He has an interest in promoting integrity in research, and has long advocated establishment of an office for research integrity in Australia.

Professor Richard Harvey was awarded a Member of the the Order of Australia (AM) for ‘significant service to medicine in the field of cell biology and cardiovascular research, and through scientific leadership roles’.

Richard was born in Adelaide and grew up in Bute, a small one-pub country town in the upper reaches of Yorke Peninsula in South Australia. He did a BSc at the University of

Adelaide, continuing with Honours and a PhD in the Department of Biochemistry under the supervision of Julian Wells.

After a short period with a biotech company in Strasbourg, France, where he cloned and expressed (in active form) the potent anti-thrombin anticoagulant Hirudin from the medicinal leech, he undertook postdoctoral studies at Harvard University with Doug Melton, where he began his life-long interest in embryonic development.

He established an independent group at the Walter and Eliza Hall Institute in Melbourne, where he spent ten years. In 1998, he relocated to the Victor Chang Cardiac Research Institute, where he is currently Co-Deputy Director and Head of the Developmental and Stem Cell Biology Division. His research focuses on the genetic basis of heart development and the pathological mechanisms underlying congenital heart disease, and cardiac regeneration. He holds the endowed Sir Peter Finley Professorship of Heart Research at the University of New South Wales, and is a Fellow of the Australian Academy of Science, Australian Academy of Health and Medical Science, EMBO and The Royal Society of London. He has been a member of the ASBMB since 1998 and in 2010 was awarded the Lemberg Medal.


Catching the Biology BugMost scientists seem to express their scientific nature

early in life, and I was no exception. I was the youngest of five children growing up in a suburb of Philadelphia. My father was a chemist at DuPont, developing specialist spray paints for General Motors cars and the lining of Coors beer cans, among other things, while my mother was a self-taught librarian and local historian. I spent most of my childhood days catching frogs in our ponds, collecting all manner of natural ‘treasures’, playing with my stuffed animals (I never really played with dolls), and roaming the neighbourhood with my friends, as you could in those days. My scientific side was fostered by my father, who was also a keen fisherman and keeper of fish – I knew the Latin names of most of his tropical killifish, including my personal favourite, Aphyosemion bivittatum (pronounced exactly as it’s written, folks). I loved animals and always thought I would make a living doing something involving animals. (Lesson 1: Your basic personality develops early in life, so revisiting your childhood can help you remember who you are.)

My childhood and adolescence were pleasant and despite my early conviction of a career centred on animals, I toyed with other options, including being a doctor and writing poetry. Mostly, I just drifted through school, although I was deemed smart enough to be in the ‘advanced placement’ group and managed good enough grades. When the time came for university, I didn’t look far – just an hour down the road (albeit in a different state) at the University of Delaware (UD). I entered UD’s fledgling Honours Program in the autumn of 1980, which meant I was in a dorm with other Honours students and got to attend smaller freshman classes of about 25–30 students instead of ten times that number. I was a Biology major, and my freshman Honours Biology class was taught by Steven Skopik, who was fabulous! He was both enlightening and entertaining, and a perfect example of how a great lecturer can ‘turn you on’ to the subject. I definitely caught the bio bug, thanks to Professor Skopik. (Lesson 2: Never underestimate the value of good teaching.)

My years at UD saw my first forays into research. During my second year, I spent a semester getting lab

experience on the role of the pineal gland in non-shivering thermogenesis in woodchucks, which was intense (especially when I killed one by accidently giving it ten times the intended dose of adrenalin – oops). Then, the next summer, I worked at the Monell Chemical Senses Center in Philadelphia investigating how alpha female marmosets (a type of little monkey) use pheromones and social cues like grooming to suppress reproduction in subordinate females. In my final year at UD, I did a thesis project (essentially the same as Australian Honours projects) on the role of the sympathetic nervous system in the vasopressor effect of angiotensin in bullfrogs (yes, frogs again!). That thesis project resulted in my first published paper in General and Comparative Endocrinology. Although these projects were disparate in scope and species, they all had one thing in common – communication between distant cells via chemical messengers, ie, endocrinology. To this day, endocrinology is still my favourite discipline. (Lesson 3: Pursue a range of topics in your research career; you will likely find that the things that interest you have a common underlying theme. Oh, and, always double-check your calculations!)

PhD YearsI did very well at UD and finished up with a 4.0 grade

point average, which meant I could pretty much pick and choose where to go for a PhD (I never considered anything but more study and research). My short list came down to Cornell University (where my dad did his PhD), with its focus on veterinary science, and the University of Virginia

Communicating the Message

Rebecca Lew recounts her journey from budding young biologist to endocrinology researcher and medical writer.

Rebecca Lew.

Atop the now-gone World Trade Center in New York with fellow PhD students, circa 1985.


(UVA), with a strong physiology department. I visited both campuses, but was swayed towards UVA mainly because the PhD students there were great fun! Unlike in Australia, most PhD programs in the US (at least in those days) take 4 to 5 years, and include a coursework component as well as the independent research project. UVA Physiology was particularly tough – two solid years of coursework before you were allowed to start your PhD project, although you were encouraged to do lab rotations during your second year to help decide on your supervisor and project. I personally think that this format provides you with an excellent, well-rounded scientific education, beyond just your focussed field of research.

I didn’t appreciate it at the time, but the physiology department at UVA in the ‘80s was among the very best in the world, with big names like Bob Berne (department chair), Brian Duling, Gary Owens, and many more who Australian biochemists have probably never heard of – you’ll just have to trust me! (Lesson 4: Appreciate and make the most of the scientific talent around you.) In part because of my interest in endocrinology, I selected Alex Baertschi as my supervisor. Alex was a Swiss import who had a number of research interests in the neuroendocrine field. I dabbled with a couple of ‘out there’ projects, which in retrospect were really cutting-edge, including investigating the neuroendocrine-immune axis, which would have been really cool if I’d been able to make it work... In the end, I settled on investigating how hypoxia stimulated the release of a newly discovered hormone from the heart, atrial natriuretic peptide (ANP), using both isolated rat hearts and cultured atrial cells.

Southern CharmWhen I decided to go to UVA, my grandmother, whose

family was originally from the South, said “You’ll meet some nice Southern gentleman who you’ll marry.” Well, it turned out the gentleman was from further south than Nana intended! Towards the end of my second year, an Aussie postdoc joined Brian Duling’s lab, next door to Alex’s lab. This guy had red hair, a bushy beard, a funny accent, and was loud and brash – not anything like what I was looking for in a boyfriend. Crocodile Dundee had just come out, so Aussies were all the rage (apparently), but

that didn’t entice me the first few times Michael asked me out. However, his persistence (and charm) eventually won out, and by the start of 1987, we were officially an item.

After a year or so, we decided to get married, which we did in April 1989. I was still about a year away from finishing my degree, so Michael had to scrounge to find money to extend his time in the States. His particular visa type required him to leave the US when his postdoctoral studies were complete, and he could not work in the US for two years after leaving – this despite being married to a US citizen and at the same time as a general amnesty for illegal immigrants was declared (Mike, of course, was legal – he’d have been better off if he weren’t). So it was always clear that I needed to find a postdoc in Australia, which in those pre-internet, pre-email days, wasn’t so easy. I started looking through the journal Endocrinology, and one name kept cropping up – John Funder. I wrote to Funder, who replied that he was going to the Endocrine Society meeting in Seattle and we could meet up there. It was a brief, but apparently successful, meeting, and eventually, I had a postdoc lined up with Ian Smith, who worked with Funder at the old Prince Henry’s Institute of Medical Research (PHIMR) off St Kilda Road in Melbourne. Michael and I arrived in Melbourne on Anzac Day 1990 (after some harrowing days waiting for my visa to be approved), and I started work at PHIMR soon after. (Lesson 5: Don’t be afraid to venture far from home, both literally and figuratively.)

Early Days with Enzymes in the Land of OzFunder had a project idea for me, which took advantage

of my background in cardiac hormone research and Ian’s expertise in high-performance liquid chromatography, which I was quickly schooled in. The idea was based on the high expression of an amidating enzyme in the atria of the heart, the same tissue that synthesised ANP. This enzyme catalyses the C-terminal amidation of many hormones – but ANP was not one of them. The hypothesis was that there was another atrial peptide hormone that was amidated – so the search began. Ian and I developed methods to fractionate and assay heart extracts in such a way as to identify putative amidated peptides. The idea was sound in theory and the techniques worked on control samples, but we never did find anything. To the

With future husband Michael in the Everglades, circa 1988.

Becky and son Brian with the Ian Smith lab on the occasion of Corie Shrimpton’s PhD at the Baker Medical Research Institute in 1998.


best of my knowledge, no one else ever has either, and the accepted wisdom now is that the amidating enzyme plays a completely different role in the heart than in other endocrine tissues. I spent just a few months at PHIMR before moving to the Baker Medical Research Institute when Funder became director there.

The Baker was, and still is, known for its focus on cardiovascular disease, and was where my husband did his PhD work. Michael had returned to the Baker after UVA, so we were reunited (for a while, anyway) with my move from PHIMR. The Smith lab settled in well, and what followed were some very enjoyable years with great colleagues. After the quest for the elusive amidated atrial peptide fizzled out, I branched out to other enzymes involved in the metabolism of peptides regulating cardiovascular function. The most famous such enzyme is angiotensin converting enzyme (ACE), the target for the antihypertensive ACE inhibitor class of drugs that are in widespread therapeutic use. However, we focussed on more obscure enzymes, particularly one affectionately known as ‘24.15’, after its Enzyme Commission number, EC 3.4.24.15. This enzyme is involved in the degradation of several bioactive peptide hormones, including the vasodilator bradykinin. In collaboration with labs in Australia and worldwide, the Smith lab characterised 24.15 and its role in peptide metabolism. I spent most of the 1990s dedicated to this little enzyme (confession: throughout the year 2015, I would often say “2415” because I had said it so often) and its cousins, but came up short on anything groundbreaking. However, I enjoyed the work, my labmates, and annual pilgrimages to the Lorne Protein Meeting, and Ian kept us well funded – what more could I want? Although the publications came at a steady pace, I suppose my most important achievements during this period were the creation of two sons, Brian in 1997 and Daniel in 2000. (Lesson 6: As much as I enjoyed the Smith lab, I realise in retrospect that I was too comfortable and spent too long there – be sure to change jobs a bit more often than every 15 years…)

To Monash and BeyondIn 2004, the Smith lab moved to the biochemistry

department at Monash, soon after Chris Mitchell took over as head. The lab had been collaborating with people there, such as Mibel Aguilar and Patrick Perlmutter, for several years, and both Ian and I had given lectures there. Thus, the ties were strong, and the department was vibrant and more aligned with where our research was heading than the Baker. During this period, I started to branch out on my own a bit more and come out from under Ian’s shadow (Ian’s a big man, both physically and in personality and reputation). I had considered leaving Ian’s lab on several earlier occasions, but circumstances and inertia prevented that, undoubtedly to the detriment of my research career. One area of interest was lecturing, so I increased my involvement with the departmental education group. Another area was academic publishing – I was one of those freaks who actually enjoyed writing papers – so when I read in the Australian Biochemist that the then-Editor, Phillip Nagley, was looking for people to join an editorial committee, I signed up. This little step turned out to have major consequences, in that it directly led to me becoming Editor at the start of 2006, which, in turn, led to my big career shift later that year. (Lesson 7: You never know where things might lead, so if an opportunity comes up that interests you, take it.)

During the early 2000s, I began to realise that I wasn’t a star researcher and was unlikely to ever become one. Worse still, I was enjoying research less and less – having to focus on one, relatively narrow topic was, quite frankly, becoming boring. After a couple of false starts with alternative careers, I stumbled upon an ad in Seek for a ‘medical writer’, a job which I had never known existed, with a company called ProScribe. I applied, completed the writing assessment and interview, and was offered the job! Now being the cautious person I am, I didn’t want to burn my bridges behind me, so for the first year or so, I worked half-time with ProScribe and half-time as a lecturer at Monash. I had great flexibility, thanks to both employers, especially as ProScribe at that time was a ‘virtual company’ with only home office-based writers. It didn’t take long before I knew I had found the perfect job for me.

ASBMB Editorial Committee meeting held at ComBio2005 (clockwise from top left): Sam Richardson, Nghia Le, Liana Friedman (Editorial Officer), Agnieszka Lichanska, Graham Baldwin, Phillip Nagley (ASBMB President standing in for

Editor Clem Robinson) and Rebecca Lew (Editor Elect).

Rebecca with ProScribe colleagues at a rope-climbingactivity at Taronga Zoo, March 2017.


Career SwitchMedical writing mainly includes assisting authors with

manuscripts, abstracts, posters, and slide presentations (usually for clinical trials), but can also include a range of other document types, such as submissions to regulatory agencies like the Therapeutic Goods Administration. Although the focus of a medical writing job is, of course, writing, there are many other aspects that I discovered I was good at. Analytical thinking, project management, attention to detail, diplomacy, efficiency, etc. The job is as deep as it is wide – everything from understanding the science behind a study and its place in the literature down to grammar and punctuation. Over the years, I have worked on a broad range of projects (including manuscripts, abstracts, posters, slides, grants, medical education programs, and regulatory documents) in a broad range of therapeutic areas (including oncology, diabetes, ophthalmology, psychiatry, dermatology, gastroenterology, neuroscience, osteoporosis, and more). I’ve also worked with countless clients and authors from around 25 countries worldwide and have had the opportunity to travel to several Asian countries as part of my job. Needless to say, boredom has never been an issue! I have also had the privilege to work alongside some incredibly smart, dedicated people, and have even helped train a few with ties to ASBMB… (Lesson 8: Understand your own interests, skills, and limitations, which will help you find a career that suits you best.)

Despite having been a medical writer for ten years and rising through the ranks, I feel like I am still a beginner and am learning all the time. This is probably the most rewarding aspect of my job – there is nothing quite like the feeling of starting a new project and knowing that, although you have done similar projects in the past, each one will bring new challenges and new rewards. I believe that continually challenging yourself is a critical part of fulfillment, whatever your career choice may be.

Rebecca with husband Michael and sons, Daniel and Brian (right).

ELECTION OF COUNCIL 2018Nominations are called for the following positions on the Council of the Australian Society for Biochemistry and Molecular Biology Inc for 2018: Secretary, Treasurer, Editor, Secretary for Sustaining Members and State Representatives for ACT, NSW, Qld, SA, Tas, Vic and WA.

President L. TilleyPresident Elect nominations sought Secretary B. Forbes#Treasurer T. Piva*Editor S. Mathivanan#Education Representative S. Rowland#Secretary for Sustaining Members S. Jay#

ACT Y.P. Mabbitt*NSW K. Michie#Vic E. Lee#Qld D. Ng*SA S. Polyak#Tas K. Brettingham-Moore#WA N. Taylor*

Nomination forms are available on the ASBMB website. Nominations for all vacant positions must be signed and seconded by members of the Society. The nominations must be signed by the nominee to indicate his/her willingness to stand. If more than one nomination is received for any position, a ballot will be held to determine the successful candidate. All members may vote for all positions except those of State Representatives where election is by members in the State concerned.

NOMINATIONS MUST REACH THE SECRETARY BY 5PM 21 SEPTEMBER 2017 (14 DAYS BEFORE THE ANNUAL GENERAL MEETING TO BE HELD AT 1:15PM 5 OCTOBER 2017)

* Retiring member, not eligible for re-election

# Eligible for re-election

The Council for the period 1 January 2017 to 31 December 2017 is composed of the following members:

Representatives for:


Sydney Protein Group:an ASBMB Special Interest Group

HistoryThe Sydney Protein Group (SPG) has a rich history of

promoting protein science locally. The SPG is made up of protein scientists and students from academia, hospitals and industry that meet regularly to hear updates from local and international speakers on the latest developments in protein science.

SPG Meetings and Early Career Researcher SupportThe SPG organises and supports several annual events,

with the help of funding from the ASBMB and dedicated trade partners.

An important Sydney Protein Group ethos is to provide opportunities for young researchers working in the field of protein science to attend high quality scientific meetings, where they can hear about recent research, interact with guest speakers and other researchers, and present their own work. An initiative of the ASBMB in recent years is to offer a SIG-sponsored speaking slot at ComBio for a promising early career researcher. This year the SPG-sponsored early career researcher who spoke at ComBio2016 was Laura McCaughey from the University of Technology Sydney (UTS). Laura talked about characterising a new protein antibiotic from Pseudomonas aeruginosa and its potential as a new target in the fight against bacterial drug resistance.

The SPG also offers several prestigious prizes aimed at PhD/honours students:

1) The annual Thompson Prize is in recognition of the eminent local protein scientist, EOP (Ted) Thompson. The Thompson Prize was inaugurated in 1992 and is awarded for the best oral presentation by a young local scientist in the field of protein structure and function.

The Thompson Prize moved further afield in 2016: it was held as a half-day meeting at the Illawarra Health and Medical Research Institute, University of Wollongong (UOW), hosted by Heath Ecroyd and Justin Yerbury. Tony Ngo (Victor Chang Cardiac Research Institute) kicked off the talks by describing his work on CoINPocket and how he has used this bioinformatics tool to predict ligands for orphan GPCRs. Emma Dawson (UTS) described her work on newly characterised dynamin-like dlp proteins and how they contributes to membrane integrity of Helicobacter pylori, suggesting a possible future drug

target. Next, Lisa Belfiore (UOW) spoke about her PhD research on the development of drug-loaded liposomes to target breast cancer cells. Her work included the use of single molecule fluorescence to quantify how many targeting moieties there were per liposome — a first of its kind! Neil Robertson (University of Sydney; USyd) told us about his innovative FRET-based techniques to elucidate the binding kinetics of the LIM-LID domains. These proteins are notoriously difficult to work with in isolation and hence were not amenable to standard biophysical analyses. Samira Aili (UTS) introduced us to the world of ant venom and its potential as a source of bioinsecticides. Her lead compound(s) looks promising and could one day be applied in the field to help ensure our food security.

As expected, the talks were of superb quality and the range of topics certainly impressed the judges [consisting of Margie Sunde (USyd), Mark Molloy (Macquarie University) and Martina Sanderson-Smith (UOW)]. In the end, the judges agreed that Emma Dawson was to be awarded the 2016 Thompson Prize.

2) The SPG also award three annual Lorne Travelling Scholarships to help the best and brightest PhD students attend the Lorne conferences.

The Lorne Travel awards went to Neil Robertson (USyd) winning the ATA Scientific Lorne Travel Award and Claudia Kielkopf (UOW) winning the SPG Lorne Travel Award. The Greg Ralston Memorial Award (Honours student continuing on to do a PhD) went to Gabrielle McClymont (USyd).

3) Lastly, look out for the East Coast Protein Meeting (ECPM) this year in Coffs Harbour (14–16 July), organised by the SPG. The ECPM is a joint initiative with the Queensland Protein Group that is held every two years. The focus of this meeting is on Early Career Protein Researchers who present their work alongside a few keynote speakers.

Jason Low and Liza Cubeddu website: www.mmb.usyd.edu.au/spg/

2016 Thompson Prize finalists and judges.From left: Margie Sunde (USyd – Judge), Lisa Belfiore

(UOW), Tony Ngo (VCCRI), Neil Robertson (USyd), Emma Dawson (UTS), Samira Aili (UTS), Martina Sanderson-Smith

(UOW – Judge) and Mark Molloy (Macquarie – Judge).

Laura McCaughey (UTS) was the SPG-sponsored

early career researcher

who spoke at ComBio2016.


The Metabolism and Molecular Medicine Special Interest Group (MMM-SIG) is one of the largest in the ASBMB, with more than 120 members. Since 2014, the group has been under the direction of Nigel Turner (University of NSW) and Sean McGee (Deakin University). The goal of the MMM-SIG is to support activities that showcase the best research in this field from Australia and abroad. This has included active contributions to the ComBio meetings, as well as financial support for other conferences that are relevant to members of the MMM-SIG.

In 2015, the MMM-SIG was a sponsor for the 1st Australian Cancer and Metabolism Meeting (ACMM 2015). This meeting was initiated to provide an opportunity for scientists with expertise in nutrient metabolism to engage with cancer biologists and discuss the latest developments and advances in this rapidly expanding field. The meeting was attended by approximately 180 delegates and the broad topics covered across the meeting were the involvement of lipid metabolism, nutrient uptake, obesity and metabolic signalling in the regulation of cancer cell growth, as well as a session on therapeutic targeting of metabolic pathways to treat cancer. The keynote speaker for the meeting was Professor Brendan Manning from the School of Public Health at Harvard University. He gave an excellent presentation on the control of nutrient metabolism and cancer cell growth by the mechanistic target of rapamycin (mTOR). Throughout the meeting, there were a number of other stimulating talks from invited and selected speakers, a dedicated session for student presentations and a lively poster session in which there was very active engagement and discussion across the 50 poster presentations. Overall, this inaugural meeting was a great success and showcased the breadth and depth of outstanding cancer metabolism research being performed by Australian researchers.

In 2016, the MMM-SIG organised a symposium on ‘Nutrient Sensing Mechanisms and Pathways’ at the ComBio meeting in Brisbane. We were lucky to attract Dr Matthew Hirschey from Duke University as an international speaker for this symposium. Dr Hirschey opened the symposium with a fantastic presentation that described how metabolites generated from the metabolism of fatty acids can directly alter histone acetylation. This represents a novel epigenetic mechanism linking dietary fuels to the reprogramming of gene expression. Associate Professor Zane Andrews from Monash University then presented new findings on the metabolic role of the enzyme carnitine acetyltransferase and how its deletion from specific hypothalamic neurons can influence whole-body glucose and lipid metabolism. The third speaker in the symposium was Professor Amanda Page from the University of Adelaide, who presented research on how

vagal afferents from the stomach control satiety and how sensitivity of these neurons changes in a diurnal fashion and in response to nutrient excess. Dr Frances Byrne from the University of NSW then gave an excellent overview of the role that a specific glucose transporter, GLUT6, plays in the development of endometrial cancer. The final speaker of the symposium (short talk selected from the abstracts) was Stephen Fairweather from the Australian National University, who discussed the physiology and biochemistry of amino acid transporters.

In 2017, the MMM-SIG will be supporting the 2nd Australian Cancer and Metabolism Meeting scheduled for 15–17 May at the Victorian Comprehensive Cancer Centre, Melbourne. This is sure to be an outstanding highlight on the conference calendar for 2017, with a plenary lecture to be delivered by one of the world leaders in this field, Professor Ralph Deberardinis from the University of Texas Southwestern Medical Center. The meeting will cover areas including immuno-metabolism and cancer, metabolic disease, lifestyle interventions and cancer, novel metabolic pathways in cancer, and cancer metabolism and therapeutics. The MMM-SIG will also continue to support the attraction of world-class speakers at the upcoming ComBio2017 meeting in Adelaide.

Given the increasing recognition of the role of aberrant metabolism in many major diseases, the MMM-SIG is committed to supporting relevant meetings in this field. While this support may not always be financial, we can assist you to enhance the quality of your meeting by connecting you with our network of enthusiastic scientists from across the country. Please feel free to contact us if you have proposals for future ComBio symposia or if we can support you in any other way.

Nigel Turner and Sean McGeeemail: [email protected]

Metabolism and Molecular Medicine: an ASBMB Special Interest Group

Enjoying the welcome mixer at ComBio 2016. From left: international speaker Matthew Hirschey (Duke University),

Arthe Raajendiran, Matthew Watt and Sean McGee.


The Victorian Branch of the ASBMB continues to be a proud sponsor of the Science Talent Search, which was held in October last year at La Trobe University. This popular annual event founded by the Science Teachers’ Association of Victoria in 1952, was established with three broad aims:1. To stimulate ongoing interest in serious study of the

sciences by encouraging independent, self-motivated project work amongst students of science, giving students the opportunity to communicate their achievements to a wider audience and recognising effort and achievement in their scientific enterprise.

2. To promote the direct involvement of the students in the process of science and its communication.

3. To give the public an opportunity to see the quality of work being achieved in science, by both primary and secondary school students.

As part of the event, students enrolled in Prep through to Year 12 are invited to submit projects in ten categories: Computer Programs, Games, Science Photography, Posters and Scientific Wall Charts, Working Models, Inventions, Experimental Research, Creative Writing, Video Productions and Class Experimental Research Project.

The theme for 2016 was ‘Drones, Droids and Robots’. There were a total of 2154 entries from 3089 student participants from 153 Victorian schools.

The Victorian Branch of the ASBMB supported the 2016 Science Talent Search with a $1,000 donation, which was awarded as major and minor bursaries to 25 students from both Victorian Primary and High Schools, including: Box Hill High School, Brighton Primary School, Eltham College, Fintona Girls School, Firbank Grammar School, Lab Rats Science Club, Plenty Valley Christian College, Shelford Girls’ Grammar, St Leonard’s College, St Margaret’s School and Wesley College.

The major Experimental Research and Inventions winners were then provided the fantastic opportunity to attend the National BHP Billiton Science and Engineering Awards at the end of 2016. Here, Victorian students were

well represented in the finals with two of our young budding scientists placing in the top three of the Inventions (Engineering) category.

We are extremely fortunate to be in the position to be able to contribute to the development of young aspiring scientists and we look forward to the opportunity to support this initiative for many years to come.

Erinna Lee, ASBMB Victorian State Representativewww.sciencevictoria.com.au/sts

Science Teachers’ Association of Victoria

Science Talent Search

Right: Images of a banana’s chromosome clumps at

resolution of 0.5 microns.

Left: A St Kevin’s College senior school student receives

the first place award in the Engineering Inventions

category of the BHP Billiton Science and Engineering Awards. His project was titled “Synth-etic: The

world’s first ‘Hood-Wind’ instrument”. He is pictured with the Science Teachers’ Association of Victoria’s President, Soula Bennett.

Opening ceremony.


So I Filed a Provisional Patent Application – What Next?

A series of regular articles on intellectual property.

In this issue, Sarah Hennebry Patent Attorney, FPA Patent Attorneys, outlines the steps

following the filing of a patent application.

IntroductionMy previous article in the December 2016 Australian

Biochemist discussed the type of information you might wish to provide to your patent attorney when you are first considering filing a patent application, or when you have engaged your attorney to draft a patent application.

This article looks at what you may need to do after your application has been filed, and what happens to your application should you decide to continue pursuing the application after 12 months.

First StepsGenerally, the first application that is filed in the patenting

process is a ‘provisional’ (sometimes called ‘basic’) application. This type of application is not examined by the patent office, and lapses after 12 months.

In some instances, it is possible to skip the step of filing the provisional application and proceed straight to filing a complete application. This, however, is not typical.

12 Months LaterIf, after 12 months from filing the provisional

application, you still wish to pursue patent protection, a ‘complete’application must be filed. • the complete application must be filed on or before

the 12 month anniversary of filing the provisional application;

• the complete application will be given a ‘priority date’ which corresponds to the date on which the provisional application was filed.

If you don’t want to pursue patent protection, or are concerned that more time is needed to develop the invention, no action is required and the provisional patent application will lapse (and the contents of the application will not be published, other than the title and applicant details).

Content of the Complete ApplicationIt is important to understand that your involvement with

the patent application does not end after the filing of the document.

The preparation of the complete application is the last opportunity for a patent application to include data and a description of how to perform the invention. After filing the provisional application, you should continue to develop the invention and perform experiments which are relevant for inclusion in the complete application.

It is increasingly difficult to obtain a claim that is broader than exactly what is shown in the examples of a patent specification. If the claims of a patent are restricted to the specific examples provided in the specification, then third parties may be able to design around a patent claim, and this may reduce the commercial value of the patent.

Some types of data can assist in broadening the scope of a patent claim. Ideally, these data should be included in the provisional application, but can still be of significant benefit if obtained during the 12 months following the filing of the provisional, and included in the complete application. Some experiments and data to consider are shown in Table 1.

Complete Applications and International Applications

The complete application can be one or more individual national applications or alternatively, an ‘international application’ (also called a ‘PCT application’). It is important to understand that there is no such thing as an ‘international patent’; an international patent application is simply a bundle of national applications and enables the applicant to file a single application and then designate a large number of countries at a later point in time (see Fig. 1).

One benefit of filing an international application is that more time is provided to obtain relevant funds for pursuing patent protection (including time to find a commercial partner, if needed). Furthermore, filing an international application buys you more time to decide on the countries in which patent protection will be sought.

The international application will be published approximately 18 months after you filed your provisional application (or 18 months after filing your international/complete application if you skipped filing a provisional

Fig. 1. Timeline of patent filing and prosecution.

Document Title Position and Style Document Title Position and Style

0 12 months 30 months

Formalities, examination, grant

up to 7-10 years

IP Development

18 months


application). This is another significant deadline to consider, particularly if you have not yet published details of your invention elsewhere. It is possible to withdraw the international or complete application prior to publication, if you want to avoid disclosing your invention to the public. Once this deadline has passed, your patent application will be in the public domain and it will no longer be possible to keep your invention secret.

Approximately 18 months after filing a PCT application (or 30 months after the provisional application was filed), it is necessary to designate the specific countries in which a patent is desired. It is not possible to obtain patent protection in all countries through a PCT application and you should discuss this with your patent attorney so that appropriate action is taken at an appropriate time.

ExaminationIn most jurisdictions, examination of the patent

application is performed before a decision is made to ‘grant’ a patent. This examination is performed by examiners employed by the national patent offices in each country. The technical background of patent examiners varies from country to country.

There is significant variation in the time taken for examination in each jurisdiction, as well as the start date of examination. While in some cases it is possible to expedite the examination process, in extreme cases it can take up to ten years after filing an application for examination to be complete. During this time, your patent attorney may require your input at different stages, to assist them in prosecuting your application through the examination process.

The examination process includes:• an assessment of the claims made in the patent

application to determine if they are eligible for patent protection under the patent laws of that country;– are the claims novel (new) and inventive (not

obvious)? – this assessment will be made by comparison with

any disclosures that are in the ‘prior art base’ (see Glossary below).

• (in most jurisdictions) an assessment of the quality of the patent specification, including– whether the specification provides sufficient

instructions for the person skilled in the art (PSA; see Glossary below) to perform (would ‘create’ or ‘recreate’ be a better word?) the invention;

– whether the skilled person was ‘in possession of the invention’ at the time of filing the application.

• if a patent examiner considers that there is a lawful reason to object to one or more claims made in the patent application, a report will be issued– the patent applicant may respond to the report,

either by amending the claims, submitting arguments or a combination of both approaches.

• if the examiner’s objections cannot be overcome within the allocated time, the patent application will lapse.

The tests for patentability that are applied in each jurisdiction vary and it is important to recognise that just because one Patent Office will grant a patent, not all Patent Offices will do the same.

It is common for patent attorneys to require the input of the inventors during the examination process. For example, the inventors’ technical expertise and understanding of the prior art may be required for the purposes of rebutting any assertions made by the patent examiners. Further, declarations made by the inventors can also be of use to clarify the state of the art prior to the invention, and for highlighting difficulties in arriving at the invention (supporting, for example, inventive step arguments).

What Happens After Examination?After examination, and once the patent examiner

considers that the claims are allowable, the patent application will proceed to ‘grant’.

In most countries it is necessary to pay periodic fees to maintain the patent application and any patent granted on that application up until expiry of the patent. The cost of these fees varies considerably between countries and are payable up until expiry of the patent. If the fees are not paid, it is possible that the patent application will lapse (or in the case of a patent, it will cease).

How Long Does a Patent Last?In most jurisdictions, the term of a patent is 20 years.

This is calculated from the filing date of the complete application. There are variations to this term, for example, where the patent is directed to certain pharmaceutical products that require registration through a regulatory body (such as the Therapeutics Goods Administration in Australia).

What Rights Do I Have Once My Patent Is Granted?

Once a patent has been granted, a patentee may sue for infringement of the patent, if they believe that there is an existing (or sometimes threatened) infringement of a patent. Because patents are granted on a national basis, infringement of a patent must also be established on a national basis, and with reference to the specific claims that are granted in each relevant jurisdiction.

SummaryIt is clear from the above that obtaining patent protection

is a lengthy process and the involvement of the inventors does not end after the filing of the initial provisional application. Obtaining patent protection requires planning and forward thinking to ensure that applicants aren’t caught out or limited to claiming a subset of their true invention. Discussing the overall process with your patent attorney in the early stages of patent drafting will help to make the process as smooth as possible.




Table 1. Data to include in a patent application.

Generate a class Try to create a group or ‘genus’ from a specific example.

80% solution This is a question of how someone might follow an inventive idea, without replicating the invention exactly and avoiding infringement.

Test drive other models Extrapolate from one context or disease type to another.

Negative results Even negative results can be informative and helpful in obtaining a patent, even if they don’t make it into the final publication in a scientific journal.

Consider the case where a single amino acid substitution in an enzyme results in increased enzymatic activity. If only one example is provided in the application (for example, substitution to a tyrosine) then the patent claims may be limited to the specific substitution.Data showing that mutation to any aromatic residue also gives rise to the altered activity, may provide a generalisation that enables the applicant to obtain a broader claim.

Inventors tend to try to solve a problem completely and perfectly. When briefing their attorney, inventors often focus on the ‘100% solution’ (as discussed in the December 2016 Australian Biochemist). Consider a small molecule inhibitor of an enzyme. Consider the core structure of the molecule and the options for substituents. Do the data show that only one variation in the molecule’s structure is useful of inhibiting enzyme function or is it possible to make certain alterations to the substituent, without significantly affecting the inhibitory function of the molecule? Considering the answers to these questions, and providing data showing that modifications to the structure do not affect function, may enable the patentee to claim a broader class of structures.

Consider the scenario where initial experimental data show efficacy of a molecule in inhibiting tumour growth and reduction in tumour size, but only in one animal model of a specific cancer. Test the molecule on related cancer animal models so that the results can be extrapolated to the use of the molecule in treating more than one type of cancer.

Although negative results typically don’t make it into journal articles, these data can be helpful in obtaining patent protection. For example, if a number of experiments were conducted but didn’t work, this may support subsequent inventive step arguments (i.e., that the solution to the problem being solved was not necessarily obvious or the skilled person did not have ‘an expectation of success’). Negative results can also be useful if trying to claim the synergistic combination of known compounds. For example, if synergism only occurs within the specific range of concentrations of the actives, then negative data can show that simply combining the known compounds would not necessarily lead to the result (again assisting in an inventive step argument).

Table 2. Patent glossary.Term Inventive step

Novelty

PCT application

Person skilled in the art (PSA)

Prior art base

Priority date

Definition A subjective test is applied to the patent claims and asks the question of whether the claims are obvious. This question is answered by considering the knowledge of the ‘person skilled in the art’ as at the priority date of the patent application. There may be variation in the way this test is applied in different jurisdictions.

A determination of whether the patent claim relates to a ‘new’ invention, by reference to publically available information, including patent literature and non-patent literature. In some jurisdictions, ‘actions’ and trade displays may also be considered when assessing novelty.

An international application that allows a patent applicant to make use of an administrative process to file the same application in multiple countries at the same time.

A hypothetical person adopted for the purposes of determining whether a patent claim involves an inventive step. This person is said to have the skills and background knowledge of someone working in a particular technical field, but does not necessarily have inventive capacity. When considering whether a claim is inventive, examiners (or Courts) will put themselves in the shoes of the PSA.

The patent and non-patent literature that is considered against the claims of a patent application. The rules for constructing the prior art base may differ between jurisdictions. For example, in some countries the disclosures of a few documents can be combined, particularly for inventive step determination.

Usually this is the date that the provisional application is filed and must be no more than 12 months earlier than the filing date of the complete application. In some cases, where no provisional application is filed, the priority date will be the same date as the filing date of the complete application.


I was fortunate to be awarded the LabGear Australia Discovery Science Award in 2015. The award is a travelling lectureship to enable the awardee to present his/her work at centres within Australia and New Zealand and to also present a symposium talk at ComBio.

Over my career, I have been lucky to be able to explore research topics in a number of different areas across cell biology and immunology. One constant in my wandering research life has been a fascination with the form and function of the Golgi apparatus, which has followed me throughout my career. The LabGear Award was for the contributions to this topic. Our historical understanding of the functions of the Golgi has traditionally been restricted to the regulation of glycosylation and membrane transport. However, I believe there are many dark secrets to the Golgi apparatus and indeed some are now beginning to be revealed. For example, advances over the past few years have provided clear evidence that the Golgi contributes to the regulation of higher order functions such as cell polarisation, directed migration, directed secretion, metabolism and autophagy. The symposium I presented at ComBio2015 in Melbourne discussed our work on these higher functions and on the emerging role of membrane tethers in regulating Golgi morphology. Our findings were also beginning to reveal some unexpected functions associated with the ribbon structure of the Golgi apparatus, which is the typical structure found in vertebrate cells. An exciting possibility is that the Golgi acts as a major sensor of the cell.

The timing of the Award was particularly apt as I was nearing the end of my ten-year stint as Head of Department (mid-2016) and would be able to focus more on research activities. My first visit funded

by the LabGear Award was to Adelaide in June 2015 to give a guest lecture to the Adelaide Protein Group (APG) and to judge the 2015 APG PhD student award. My talk focused on our work on the trafficking of the b-secretase BACE1, and amyloid precursor protein (APP) and the production of amyloid beta in Alzheimer’s disease. It is

now clear from many studies incorporating genetics, cell biology and animal models, that dysfunction in membrane trafficking events associated with endosomes and the Golgi underpin the development of a number of neurological diseases. The work by Cheryl Chia, Wei Hong Toh and Anson Tan in my lab have helped to map the intracellular trafficking pathways of both BACE1 and APP and define where the two membrane proteins converge to promote APP processing and amyloid beta production. Both Cheryl and Wei Hong have previously been awarded ASBMB Fellowships for their achievements.

The PhD student presentations at the APG were of a very high standard. The three students gave excellent talks: Heidi Neubauer (Centre for Cancer Biology), on the oncogenic role of sphingosine kinase 2; Shee-Chee Ong (Flinders University) on novel insulin analogues with therapeutic benefits; and Stephanie Begg (University of Adelaide) on the metalloproteome of Streptococcus pneumoniae. The session was held at the attractive Adelaide University Graduates Clubhouse, and hosted in a very engaging manner by Mark Corbett, Neurogenetics Research Program, University of Adelaide.

My second visit sponsored by the LabGear Award was to the Institute for Molecular Bioscience (IMB), University of Queensland, in July/August 2016 where I spent a very enjoyable and productive two day visit hosted by Rohan Teasdale and Jenny Stow. Here I gave a seminar titled ‘The Golgi apparatus: a regulator of higher order cell functions?’ where I presented work from Prajakta Gosavi in my lab, demonstrating that the Golgi ribbon structure controls signalling pathways and autophagy. Over the two days, I had very lively and stimulating discussions with Jenny and Rohan and a number of other cell biology colleagues including Alpha Yap, Rob Parton, Brett Collins, Dominic Ng (School of Biomedical Sciences) and Fred Meunier (Queensland Brain Institute). It was very exciting to hear about the latest developments at UQ and to discuss with Jenny the lattice light sheet images of macrophage ruffling she had captured on her visit to Janelia Research Campus, Virginia, USA.

I would like to thank LabGear for their generosity in providing funding for this award. It allowed many productive interactions across states with PhD students, early career researchers and senior colleagues.

Professor Paul Gleeson, Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne.

On the Road with Intracellular Trafficking2015 LabGear Australia Discovery Science Award

Paul Gleeson receives the 2015 ASBMB LabGear Australia Discovery

Science Award from Derek Brown, LabGear Australia.

Guest lecture by Paul Gleesonto the Adelaide Protein Group.

Photo: Houng Taing.

Super-resolution image of the Golgi ribbon structure stained with cis-

and trans-Golgi markers, by Prajakta Gosavi.


Professor Feng Shao receiving the FAOBMB Award for Research Excellence (including certificate and trophy)

from Professor Kiyoshi Fukui (FAOBMB President), with Professor Zengyi Chang (FAOBMB President-Elect) at left.

Paul Gleeson, FAOBMB Representative from Australia, reports on the FAOBMB Conference and Council Meeting held in Manila, the Philippines, during December 2016.

FAOBMB annual conferenceheld in the Philippines

The 25th FAOBMB International Conference and 43rd Annual Meeting of the Philippine Society of Biochemistry and Molecular Biology (PSBMB) was held in Manila at the Philippine International Conference Centre from 5–7 December, 2016. The convenor was Professor Gracia Fe Yu from the University of the Philippines, and the meeting’s theme was focused on Biochemistry and Molecular Biology in Health and Wellness, with an emphasis on translation of natural products.

The local organising committee, comprising Professor Gracia Fe Yu, Dr Apolinario Yambot (President of PSBMB) and Dr John Donnie Ramos (Immediate Past President of PSBMB) delivered a lively and highly interactive meeting. There were more than 575 participants, including 60 foreign delegates, of which there were nine plenary speakers. Particularly notable were the vibrant interactions between students and international delegates, and the organisers should be congratulated for facilitating sessions to maximise student involvement.

The meeting commenced with welcome addresses from the FAOBMB President Professor Kiyoshi Fukui (Japan), the FAOBMB President-Elect, Professor Zengyi Chang (China) and the IUBMB President-Elect Professor Andrew Wang (Taipei, China). All three highlighted the importance of the regional alliances promoted by the FAOBMB and enthusiastically promoted further expansion of the organisation.

Highlights from the scientific program included the nine plenary lectures and Award presentations. These

included a presentation on the history of the discovery of conotoxins in the Philippines by Dr Lourdes Cruz; an exhilarating presentation by Dr Danilo Tagle (NIH) on the advances in microfabricated devices to engineer ‘organs on chips’ for drug testing, a field which has now progressed to include multiple organs on a chip to analyse physiological systems; and an elegant presentation by Professor Ricky Johnson (Peter MacCallum Institute, Melbourne) on epigenetic regulation and the potential of HDAC inhibitors for treatment of leukemia.

The winner of the 2016 FAOBMB Award for Research Excellence was Professor Feng Shao (National Institute of Biological Sciences, Beijing, China) who gave an outstanding plenary lecture on host–pathogen interactions, in which he described his contribution to unravelling the mechanisms underlying the necrotic cell death pathway, pyroptosis, and the major role of this pathway in innate inflammatory responses against bacteria. He described his very elegant approaches to define the intracellular sensors and effector machinery associated with pyroptosis. His studies have also revealed the unexpected finding that pyroptosis may well be the underlying mechanism for the non-classical secretion of IL-1b.

The winner of the 2016 FAOBMB Education Award was Professor Rongwu Yang from Nanjing University, China. In his lecture, Professor Yang presented his personal experience on teaching biochemistry with his very enthusiastic style. His vibrant presentation stressed the importance of a passionate approach to teaching and he highlighted multiple mechanisms for engagement with students during lectures. He also presented examples of a range of creative prompts he uses to promote visual understanding of the concepts of biochemistry. Professor Yang has kindly consented to his talk being made available at the following link: http://faobmb.com/2017/01/09/iubmb-education-symposium-at-25th-faobmb-conference-held-in-manila-in-december-2016/

Professor Gracia Fe Yu in formal Filipino attire at the opening of the Conference.

Chinese colleagues at the Conference. From left: Professor Cong-Zhao Zhou (delegate of CSBMB), Professor Rongwu

Yang (winner of FAOBMB Education Award), Professor Feng Shao (winner of FAOBMB Award for Research Excellence) and

Professor Zengyi Chang (incoming FAOBMB President).

http://faobmb.com/2017/01/09/iubmb-education-symposium-at-25th-faobmb-conference-held-in-manila-in-december-2016/




There were other excellent talks in the Education Symposium, including those given by Associate Professor Susan Rowland (Australia) and Professor Hoon Eng Khoo (Singapore); their presentations can also be found on the link above.

Dr Lahiru Gangoda (Department of Biochemistry and Genetics, La Trobe University) was one of eight Young Scientists who were successful in obtaining FAOBMB Travel Fellowships to attend the conference and to deliver oral presentations of their work. A unique feature of the program was the special young scientist forum, in which outstanding Filipino undergraduate research students presented their laboratory-based research projects. Five finalists were selected from over 30 applications for oral presentations in a very well attended session and all talks were of a very high standard. The international FAOBMB judges, including Wayne Patrick (New Zealand) and Paul Gleeson (Australia), awarded the best presentation to Arman Ali Ghodsinia (National Institute of Molecular Biology and Biotechnology, University of the Philippines) for his research on PI3 kinase mutations and oncogenesis.

The FAOBMB Council meeting was held prior to the

conference program (Sunday 4 December, 2016), with Paul Gleeson as the Australian delegate (ASBMB representative). The meeting was attended by delegates from 15 of the 21 constituent member countries and was chaired by the FAOBMB President, Professor Kiyoshi Fukui (Japan) (2014–2016) and the Secretary-General, Professor Phillip Nagley (Australia). The President highlighted in his report the initiatives by the FAOBMB, particularly in education and in strengthening the relationship with IUBMB. Three FAOBMB members are now serving on the Executive Committee of IUBMB, namely Professor Andrew Wang (Taipei, China) as President-Elect of IUBMB, Professor Janet Macaulay (Australia) with the portfolio of education and training and Dr Avadhesha Surolia (India) as member for publications. Professor Zengyi Chang, President-Elect for FAOBMB at the time of the Council meeting, took over as President from January 2017. Paul Gleeson indicated the considerable enthusiasm of the ASBMB to bid for the IUBMB congress in Melbourne in 2024 and the FAOBMB Council was supportive of this potential bid.

The 26th FAOBMB Conference will be held in Kobe, Japan, 6–9 December 2017, as a joint meeting with a large consortium of biological science societies (ConBio2017). The next triennial IUBMB Congress will be held as a joint meeting with the FAOBMB Congress in Seoul, Korea, 4–9 June 2018. In 2019, the 27th FAOBMB Conference will be held in Malaysia.

Thanks to Phillip Nagley for his input and some of the photographs.

FAOBMB annual conferenceheld in the Philippines

Members of FAOBMB Executive Committee, Delegates to FAOBMB Council and observers at the Council meeting held in Manila, Philippines.

Paul Gleeson with a group of young Filipino scientists.

Traditional Filipino dancers at the cultural

evening of the Conference.


OUR SUSTAINING MEMBERS

DisclaimerThe Australian Biochemist is published by the Australian Society for Biochemistry and Molecular Biology Inc. The opinions expressed in this magazine do not necessarily represent the views of the Australian Society for Biochemistry and Molecular Biology Inc.

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VWR International Pty LtdTel: 1300 727 696Fax: 1300 135 123Email: [email protected] Web: au.vwr.com

Chirascan Circular Dichroism Spectrometers

Chirascan sets new standards for steady-state circular dichroism spectroscopy. It incorporates innovative optical design features to maximise light throughput, particularly in the far-UV wavelength region, and a sophisticated digital data acquisition system that facilitates the rapid collection of more accurate and precise CD spectra. Chirascan’s digital CD spectra acquisition approach ensures that unmodified CD spectra are collected and any post-acquisition smoothing of the CD spectra will be non-distorting and completely reversible. This approach also simplifies the operation – Chirascan is as straightforward to use as a single-beam spectrophotometer.

Other key features include:• Able to collect thermal denaturation

CD spectra in a single experiment; enabling identification of the secondary structural changes associated with each phase transition

• 5 detection channels: CD, Absorbance/Transmission, HT, Temperature and Voltage. Simultaneous multi-channel data acquisition ensures that all key information is recorded with every measurement you make

• Very low nitrogen usage. Rapid and efficient nitrogen purging combined with a sealed monochromator housing ensures that just 5 l/min is required for far-UV work

• Moveable detector. The detector position is easily adjustable and can be set close to the cell to optimised performance with highly scattering samples e.g. membrane proteins

A large range of accessories are available, ensuring the researcher can be confident of a highly effective and future-proof spectrometer that can be adapted as research interests evolve. These are best explored with the comprehensive brochure available on request.

Chirascan™-plus ACD is the world’s first and only truly automated CD spectrometer. Unattended operation increases the productivity of CD measurements markedly, with up to 200 samples per day now routinely achievable. The improved productivity transforms the number and type of experiment that can be undertaken and addresses key needs in formulation and bio-comparability applications.

‘qCD’ (quantitative CD) reflects a number novel capabilities that improve performance and enable absolute CD measurements (a first in CD spectrometry).

For further information contact:-Scientex Pty Ltd Tel 03 9899 6100Fax 03 9899 6122Email [email protected]

Celase® GMP Enzyme LaunchCelase® GMP is a proprietary

enzyme containing a unique blend of collagenase, neutral protease and buffer salts that are produced using avian and mammalian tissue-free raw materials, aseptically processed, sterile filtered and highly purified under GMP guidelines.

Manufactured by Cytori Therapeutics, this product line is ideal for cell isolation studies for laboratories aiming to facilitate a smooth transition from bench and animal research to downstream clinical applications.

A single, sterile, ready-to-use enzyme containing both collagenase and neutral protease is ideal for a wide range of adipose stem cell, biomedical and bioprocessing applications.

Not all research applications require the use of a GMP grade enzyme in early phase studies. However, the recent FDA guidance issued for tissue and cell products specifically cites that GMP grade reagents should be utilized in drug-type validated processes. Subsequently, both regenerative



DAINTREEs c i e n t i f i c

AUSTRALIANew Products to the DAINTREE Range from Coyote Bioscience

Coyote’s products are affordable solutions for any laboratory.

For field work or in laboratories where power sockets are sparse, the G10 and G20 Cordless Tissue Grinders from Coyote can work cordlessly for up to 10 hours. The G50 corded Tissue Grinder is adjustable from 3K to 8K rpm.

The T-heater Slim PCR Cycler and Mini8 Real-Time PCR Cycler can both be used in the field or at your bench and efficiently run on 12V or 240V. The cyclers are compatible with any commercial SYBR or TaqMan qPCR kits.

The MD-Box-Lab is a complete solution for mobile molecular diagnostics – comprising mobile molecular diagnostic devices, consumables and protective suit all in one. Like the PCR cyclers from Coyote, the MD-Box-Lab is portable and uses 12V DC power, compatible with a car charger or 240V.

The Pad3-H heating bath is a compact dry bath/chiller for up to 48 x 0.2mL tubes with precise temperature control from ambient to 100oC. Blocks are available for 0.2mL, 0.5mL, 1.5mL and 2.0mL tubes.

The H203-PROIII Chilling/Heating bath is a small programmable dry bath/chiller for up to 96 x 0.2mL tubes, but is also suitable for 5mL, 10mL, 15mL, 50mL, and 0.5mL tubes. These blocks can be mixed and matched making this a versatile cost effective solution in the laboratory.

The BATT3 Battery Pack can be used with Coyote’s products for up to 4 hours, ideal for field work.

For information please contactMoina MacaskillDaintree Scientific AustraliaPhone (03) 6376 3335Email [email protected]

compatible with widely used analytical software (e.g., UltraScan, SEDFIT/SEDPHAT, SEDANAL, DCDT+), and the same cells and rotors used with the ProteomeLab XL-A/XL-I, the Optima AUC surpasses many features of its forerunner.

LI-COR Biosciences is a leading innovator in systems for plant research, gas analysis, drug discovery, protein research, and small animal imaging. From the first low-cost light sensor filtered for the waveband absorbed by plants, to pioneering the development of near-infrared fluorescence detection systems for image-guided cancer surgery, LI-COR provides innovative solutions for researchers.

Building on this legacy of innovation, the LI-6800 Portable Photosynthesis System delivers a new experience, along with unprecedented capabilities for measuring gas exchange and fluorescence. Improved gas analyzer precision, light source uniformity, flow path design, environmental controls – and many more design features – are coupled with a touch screen interface that provides real-time guidance from the instrument.

The LI-6800 sets a new standard for gas exchange and fluorescence measurements. A user experience built from the latest technological innovations – combined with integrated game-changing improvements in performance – yield the most advanced system on the market. The LI-6800 will unlock the secrets of photosynthesis for researchers around the world.

These researchers use LI-COR technology to advance discovery and improve lives. From studies ranging from global climate change to cancer research, scientists rely on LI-COR products and services to address important biological and environmental challenges. More information at www.licor.com.

Advances in Unique Protein and Macromolecule Analysis Technology

AUC is a unique technique that enables molecules to float freely and unbound so they can be characterized in their native state. Beckman Coulter Life Sciences returns analytical ultracentrifugation (AUC) to the centre of protein research and macromolecule characterization with the next-generation Optima AUC. This method is useful for a range of applications, from determining protein molecular weight and binding efficiencies, to characterizing nanoparticle size and payload. Because its optical system enables precise analysis at up to 20 discrete wavelengths, the Optima AUC makes it easier for researchers to study complex systems, and to determine formulation, heterogeneity, molecular weight and binding efficiencies in a single experiment—and in less time than with earlier AUC technology.

With faster scanning rates, increased wavelength precision and enhanced data resolution the new Optima AUC can provide more accurate answers from each run. At rotor speeds up to 60,000 RPM, data acquisition rates on the Optima AUC are almost five times faster than the ProteomeLab XL-A/XL-I, with three times higher radial resolution, so it can generate nearly 15 times more data. The Optima AUC currently provides absorbance and/or interference optical systems, with the option to install up to three independent, simultaneous detection systems.

All system optics are contained outside the rotor chamber, making the Optima AUC easier to clean. A 15-inch (38 cm) touchscreen display indicates the progression of experiments, while remote monitoring lets researchers set up, monitor and extract experiment data from virtually any location. Though



LATEX Bead Conjugation KitsInnova Biosciences has introduced

a range of LATEX Bead Conjugation Kits. These simple to use, one-step kits for covalently conjugating antibodies, proteins and peptides (or any other biomolecule with an amine group) to specially treated latex beads without the need for extensive optimisation.

The latex conjugation reaction has been developed using Innova Biosicences expertise in simple and quick one-step antibody conjugations such as our InnovaCoat® GOLD kits (covalent conjugation to gold nanoparticles) and Lightning-Link® kits (covalent conjugation of antibodies, proteins and peptides to enzymes and fluorophores) to produce a kit unlike any latex bead conjugation product available.

Quick and easy to use:• 30 seconds to set up the one-step

conjugation reaction• 3 minutes hands-on time and

35 minutes total time until the conjugates are ready to use

Specially treated latex is resistant to aggregation:• High yields of functional conjugates

can be made without the need for harsh resuspension methods like sonication and vortexing

Only two buffers are used to test for optimal activity:• No extensive pH optimisations as

is typically required for traditional passive conjugation methods

Choice of bead colour and kit sizes:• Available 400nm latex beads in red,

blue or black• Two kit sizes available: Mini kits

are ideal for antibody screening or ‘proof of principle’ experiments, and Midi kits which are 10 times the size of the Mini kits. Bulk material is also available for further scale up

BioNovus Life SciencesPh: (02) 9484-0931Email: [email protected]: www.bionovuslifesciences.com.au

GeneWorks will be exhibiting at ComBio 2017 in Adelaide.

GeneWorkscustomerservice@geneworks.com.auwww.geneworks.com.auFreeCall 1800 882 555Phone 08 8159 6250

Selecting the Right Gel Documentation System

Ease of use and reliability have always been the biggest considerations in choosing a gel documentation system. Recently, gel documentation systems with alternatives to UV have become increasingly popular and required as researchers shift their preferences to the use of safer fluorescent dyes instead of toxic ethidium bromide. The use of alternative light sources for excitation, such as blue light, also serves to reduce the risk of UV exposure.

Axygen® Gel Documentation systems are easy to use and reliable for the fast gel imaging operations required by researchers today. It deploys a high resolution 5.4 MP camera which captures publication quality images and features an Auto Exposure tool which enables the calculation of the optimal exposure time with just a single click.

Axygen® Gel Documentation systems also feature a Darkroom control which allows the selection from UV 302, UV 365, Epi White, or Epi Blue light sources, as well as an optional Trans White light illumination tray. With many added features and options available in the market today, selecting the right gel documentation system is important. Axygen® Gel Documentation systems assist the user to in capturing hassle-free images in the least amount of time. Please contact us on [email protected]

GeneWorks is a proudly independent supplier of molecular and cell biology products and associated services based in Adelaide. Our mission is to facilitate access to innovative technology solutions and services to enable successful research or commercial outcomes for our clients.

New introductions to GeneWorks’ product range include Alvéole’s PRIMO, the world’s first multi-protein patterning system. PRIMO allows researchers to study the influence of the microenvironment on intracellular and intercellular mechanisms. We now also supply products from BRANDEL, including a novel Gradient Fractionator system used for the separation of organelles, RNA, DNA and the study of protein interactions. BRANDEL also make systems for cell harvesting and tissue perfusion.

Organisations looking for workflow automation should consider unique instruments from Art Robbins Instruments (ARI) such as the Cobra non-contact nanoliter-capable dispensers, and the Scorpion Screen Builder high speed single channel pipettor.

Excitement continues to build around Clearbridge Biomedics’ ClearCell system for viable CTC isolation, and Bioo Scientific’s NextPrep-Mag cfDNA Isolation and NEXTflex NGS Library preparation kits and barcodes.

GeneWorks’ expanding range of services include custom Genotyping (including assay design), Gene Expression Analysis and Sample Preparation which we provide to researchers and companies wishing to outsource their wet lab work. GeneWorks also provides expert Instrumentation Services to support all the equipment we supply.


SSI Vertex Pipette Tips LabGear Australia are pleased to

announce its appointment as the new exclusive distributor for the SSI Vertex range of pipette tips. The range comes in filtered and non-filtered versions and are offered with the NoStick® resin formulation as standard which enables viscous liquids to be dispensed completely. All tips are certified free from RNase, DNase, RNA, DNA, and PCR inhibitors and are manufactured from US FDA approved medical grade virgin polypropylene. LabGear also offers the full range of SSI Ultraflux PCR products including PCR tubes, strip tubes, caps, PCR plates, and sealing film. Completing the offering are CryoFreeze tubes, racks and screw cap tubes.

GC BiotechLabGear Australia are the exclusive

Australian distributors for GC Biotech. GC Biotech manufactures a range of magnetic bead based products for genomics, Sanger and Next Generation sequencing applications. The products are: CleanPCR, CleanNGS, CleanDTR clean up kits, Clean Circulating DNA from plasma or serum, CleanNA DNA and RNA isolation kits from a wide range of sample and tissue types like, blood and tissue, plant, pathogen, FFPE and plasmids.

CleanNGS delivers efficient PCR and Next Generation Library Prep clean up whilst CleanDTR offers efficient PCR and Sanger sequencing reaction clean up based on the unique paramagnetic bead technology. The Clean Circulating DNA Kit has significant uses in research, Cancer Diagnostics, prenatal screening (NIPT), Organ transplantation and Post Trauma Monitoring to produce scalable, isolation of high quality and high recovery of targeted DNA fragments. This innovative magnetic bead technology can be used in a manual setting in 96 well plates or on your current automated liquid handling

platform of the known brands available today. Seamless transition to using the CleanNA PCR clean up and isolation kits on your existing automated liquid handling platforms mean less time for optimisation of the protocol and reduced cost of reagents.

For your cost effective, automation friendly, high quality sequencing results, efficient purification of PCR products and Size Exclusion requirements for Next Generation Sequencing Library Preps, please contact LabGear Australia on 1800 LabGear (1800 522 432) or email [email protected]

UNcle and HUNK – the Perfect Pair for Studying Protein Aggregation and Stability

Do you need to rank your constructs or optimise your formulations? AXT, partnering with Unchained Labs, can help you do this through either temperature-induced stress, disrupting the 3D protein structure, or through chemical denaturation, determining the Gibbs free energy (DG) required for protein unfolding.

The UNcle stability platform uses fluorescence, static light scattering (SLS) and dynamic light scattering (DLS) to characterise protein stability. With temperature control from 15–95oC and sealed samples, you can even do long term studies to fully understand protein stability. Measurements include thermal melting, thermal aggregation, isothermal stability, thermal recovery, sizing, polydispersity and viscosity, as well as determination of the diffusion interaction parameter and second virial coefficient to help predict colloidal stability and likelihood of aggregation and protein-protein interactions.

UNcle can also measure DG – prep your samples offline with denaturants and incubate at ambient temperature and then read them on the UNcle, where

your data is analysed automatically at the end.

HUNK fully automates the process of chemical denaturation and analyses the data giving you DGs for two-state and three-state proteins. Just add your protein, formulations and denaturants, and the HUNK performs the liquid handling and intrinsic fluorescence detection to create and measure your chemical denaturation curve. You can measure up to 96 different DGs, unattended.

UNcle and HUNK have your stability and aggregation studies covered.

For more information, visit www.axt.com.au or email [email protected]

Isolab Laborgerate GmbH is a global laboratory supplier company that specialises in manufacturing laboratory consumables.

Isolab sterile production plants are well equipped (Class 10,000 sterile) clean rooms (validated by international DIN and ISO standards) with HEPA filters and positive pressure areas which are supplied with air locked doors. Isolab certified plastics are DNase, RNase and pyrogen-free, and BioClean certificated.

Some of our life science products are: sterile tubes (centrifuge, micro, cryo, PCR, K3 EDTA), caps, flasks, plates, racks, swabs, boxes, pestles, slides, petri dishes, pipettes and workstations.

Our ‘one stop shopping for laboratories’ catalogue includes: glassware, consumables, equipment, instruments and chemicals.

Catalogue: http://catalog.isolab.de/ 2016/index.html

Please see us: www.isolab.de




Recombinant Antibodies: Highly Reproducible with Tailored Specificity

Making an antibody using recombinant technology offers a number of benefits around consistency, specificity, and scalability. Recombinant antibodies are manufactured by cloning the immune-specific heavy and light antibody chains into a high-yield mammalian expression vector. The

FORTHCOMING MEETINGS

ComBio20172–5 October 2017Combined ASBMB, ASPS and ANZSCDB Annual MeetingsAdelaide Convention Centre, Adelaide

Early Registration and Abstract Deadline: Friday, 30 June 2017

ComBio2017 will be held at the new state of the art Adelaide Convention Centre located in the heart of the city on the Torrens River, and overlooking the magnificent Adelaide Oval Precinct. The Convention Centre and hotels are located a stone’s throw from the many restaurants and cultural activities that make Adelaide such an engaging and enchanting destination.

The keynote speaker for this conference is Emmanuelle Charpentier of the Max Planck Institute for Infection Biology, Berlin. Professor Charpentier is the co-inventor of CRISP-Cas9. The program will feature a number of other overseas plenary presentations from some of the best international scientists together with a number of society speciality lectures. Several poster sessions are also planned. The scientific program of the conference will include the themes:

New Horizons in Biochemistry & Molecular Biology Education5–8 September 2017Rehovot, IsraelThis education conference is supported by IUBMB and FEBS. The conference proceeds the 42nd FEBS Congress, 10–14 September in Jerusalem, Israel.Further information:Email: [email protected]: www.weizmann.ac.il/conferences/NHBMB2017

26th FAOBMB Conference6–9 December 2017Kobe, JapanThe 26th FAOBMB Conference will be held as a combined meeting with the Japanese Consortium of Biological Sciences (ConBio 2017). FAOBMB Travel Fellowships will be available for young scientists to attend this conference.Further information:Email: [email protected]: www.aeplan.co.jp/conbio2017

24th IUBMB–15th FAOBMB Congress4–9 June 2018Seoul, KoreaThis is a triennial Congress of IUBMB, combined with the FAOBMB Congress when held in our region. A Young Scientist Program will be held in conjunction with the Congress, 2–4 June 2018.Further information:Email: [email protected]: www.iubmb2018.org

• Plant Biology• Biotechnology and Sustainable

Futures• Developmental, Stem Cell and

Regenerative Biology• Proteins and Proteomics• Genomes, Epigenetics and

Bioinformatics• Cell Biology• Cell Signalling and Metabolism

Further information:www.combio.org.au/combio2017

Conference Chair:Michael [email protected]

Registration/ExhibitionSally [email protected]

resulting vectors are introduced into expression hosts (e.g. bacteria, yeast or mammalian) for the manufacturing of high-quality functional antibodies.

Recombinant antibodies offer several advantages over both traditional monoclonal and polyclonal antibodies:• Improved consistency and

reproducibility• Improved sensitivity and

specificity• Ease of scalability• Animal-free high-throughput

production

In order to provide antibodies that have excellent sensitivity with

the highest degree of consistency, Abcam have engineered recombinant versions of our RabMAb® rabbit monoclonal antibodies. We’ve combined the advantages of our RabMAb rabbit monoclonals with recombinant technology to deliver industry-leading antibodies.

Recombinant RabMAb antibodies are validated in key applications (WB, IHC, ICC/IF, IP, and flow cytometry) and species (human, mouse, and rat). They are affinity purified and selected antibodies are Knockout validated via CRISPR technology. All products are sold with our Abpromise guarantee. Visit our website www.abcam.com to search for over 10,000 recombinant RabMAb antibodies.


DIRECTORYCOUNCIL FOR 2017PRESIDENTProfessor Leann TilleyDepartment of Biochemistry and Molecular BiologyUniversity of MelbournePARKVILLE VIC 3010Ph (03) 8344 2227Email: [email protected] PRESIDENTProfessor Michael RyanDepartment of Biochemistry and Molecular BiologyMonash UniversityCLAYTON VIC 3800Ph (03) 9902 4909Email: [email protected] Professor Terrence PivaSchool of Medical SciencesRMIT University, PO Box 71BUNDOORA VIC 3083Ph (03) 9925 6503Email: [email protected] Professor Briony ForbesMedicinal BiochemistryFlinders UniversityBEDFORD PARK SA 5042Ph (08) 8204 4221Email: [email protected] Suresh MathivananLa Trobe Institute for Molecular Science Department of BiochemistryLa Trobe UniversityBUNDOORA VIC 3086Ph (03) 9479 2565Email: [email protected] DIRECTORDr Suresh MathivananLa Trobe Institute for Molecular Science Department of BiochemistryLa Trobe UniversityBUNDOORA VIC 3086Ph (03) 9479 2565Email: [email protected] REPRESENTATIVEAssociate Professor Susan RowlandInstitute for Teaching and Learning Innovation (ITaLI)University of QueenslandST LUCIA QLD 4072Ph (07) 3365 3089Email: [email protected] REPRESENTATIVEProfessor Paul GleesonDepartment of Biochemistry and Molecular BiologyUniversity of MelbournePARKVILLE VIC 3010Ph (03) 8344 2354Email: [email protected] FOR SUSTAINING MEMBERSSally Jayc/- ASBMB National OfficePO Box 2331KENT TOWN SA 5071Ph (08) 8362 0009Fax (08) 8362 0009Email: [email protected]

STATE REPRESENTATIVESAUSTRALIAN CAPITAL TERRITORYDr Peter MabbittANU College of Physical and Mathematical SciencesAustralian National UniversityCANBERRA ACT 0200Email: [email protected]

NEW SOUTH WALESDr Katharine Michie School of PhysicsUniversity of New South WalesSYDNEY NSW 2052 Ph (02) 9385 4587Email: [email protected]

QUEENSLANDDr Dominic Chi Hiung NgSchool of Biomedical SciencesUniversity of QueenslandST LUCIA QUEENSLAND 4072Ph (07) 3365 3077Email: [email protected]

SOUTH AUSTRALIADr Steven Polyak School of Molecular and Biomedical Science University of AdelaideADELAIDE SA 5005Ph (08) 8313 6042Email: [email protected]

TASMANIADr Kate Brettingham-MooreSchool of MedicineUniversity of TasmaniaHOBART TAS 7008Ph (03) 6226 4609Email: [email protected]

VICTORIADr Erinna LeeOlivia Newton-John Cancer Research Institute145 Studley RdHEIDELBERG VIC 3084Ph (03) 9496 5726Email: [email protected]

WESTERN AUSTRALIAAssociate Professor Nicolas TaylorARC Centre of Excellence in Plant Energy BiologyUniversity of Western AustraliaCRAWLEY WA 6009Ph (08) 6488 7005Email: [email protected]

ASBMB NATIONAL OFFICEPO Box 2331KENT TOWN SA 5071Ph (08) 8362 0009Fax (08) 8362 0009Email: [email protected]://www.asbmb.org.au

SPECIAL INTEREST GROUPSADELAIDE PROTEIN GROUPContact: Dr Christopher McDevittResearch Centre for Infectious DiseasesUniversity of AdelaideADELAIDE SA 5005Ph (08) 8313 0413Email: [email protected]

AUSTRALIAN YEAST GROUPChair: Dr Alan MunnGriffith University Gold CoastPMB 50, Gold Coast Mail CentreSOUTHPORT QLD 9726Ph (07) 07 5552 9307Email: [email protected]

BIOCHEMICAL EDUCATIONChair: Associate Professor Susan Rowland School of Chemistry and Molecular BiosciencesUniversity of QueenslandST LUCIA QLD 4072Ph: (07) 3365 4615Email: [email protected]

MELBOURNE PROTEIN GROUPPresident: Dr Douglas FairlieOlivia Newton John Cancer Research InstituteAustin HospitalHEIDELBERG VIC 3084Email: [email protected]

METABOLISM AND MOLECULARMEDICINE GROUPChair: Dr Nigel TurnerSchool of Medical ScienceUniversity of New South WalesKENSINGTON NSW 2052Ph (02) 9385 2548Email: [email protected]

QUEENSLAND PROTEIN GROUPChair: Dr Brett CollinsInstitute for Molecular BioscienceUniversity of QLD, ST LUCIA QLD 4072Ph (07) 3346 2043Email: [email protected]

RNA NETWORK AUSTRALASIAChair: Dr Archa FoxHarry Perkins Institute of Medical Research6 Verdun StreetNEDLANDS WA 6009Ph (08) 6151 0762Email: [email protected]

SYDNEY PROTEIN GROUPChair: Dr Liza CubedduSchool of Science and Health, University of Western Sydney, PENRITH NSW 2751Ph (02) 4620 3343Email: [email protected]

COPY DEADLINE FOR NEXT ISSUE:Monday, 12 June 2017

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