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Poster No.
Department Presenting Author
Designation Abstract Title Authors
Phd-1 Anatomy Dr. Balpreet Kaur
PhD Student Ameliorative role of Curcumin on basal forebrain of mice subjected to arsenic trioxide exposure
Kaur Balpreet, Mehta Kamakshi, Pandey Kumar Kamlesh and Dhar Pushpa
Phd-2 Anatomy
Shubhi Saini,
Deptt. of
Anatomy
Phd. Student
MATURATION OF
NEURONS AND
ASTROGLIA IN
DEVELOPING HUMAN
COCHLEAR NUCLEUS
Saini Shubhi1,
Jacob George
Tony1, Thakar
Alok2, Roy Kumar
Kallol3, Roy
Sankar Tara1
Phd-3 Anatomy Surabhi Gautam
PhD Student Yoga reduces disease activity and severity of comorbid depression and improves quality of life in Rheumatoid arthritis: A randomized controlled trial
Gautam Surabhi, Kumar Uma, Kanga Uma, Dada Rima
Phd-4 Anatomy Tolahunase Rajaneesh Madhuri
PhD Student Impact of Yoga- and Meditation-based lifestyle intervention on depression, quality of life, and cellular aging in infertile couples
Madhuri Tolahunase, Rajesh Sagar, Rima Dada
Phd-5 Anatomy Shilpa Bisht PhD Student Impact of yoga based lifestyle intervention on sperm oxidative DNA damage: study on fathers of non-familial sporadic heritable retinoblastoma patients
Shilpa Bisht, Bhavna Chawla, Rima Dada
Phd-6 Anatomy Vidhu Dhawan PhD Student Sperm transcript signature in early pregnancy loss
Vidhu Dhawan, Neena Malhotra, Nita Singh, Vatsala Dhadhwal, Rima Dada
Phd-7 Anatomy Charanjeet Kaur
PhD Student Age-related human spiral ganglion neuronal loss assessed by unbiased stereology
Kaur Charanjeet, Jacob Tony George, Nag Tapas Chandra, Thakar Alok, Bhardwaj Dayanand, Roy Tara Sankar
Phd-8 Anatomy Meenakshi Maurya
PhD Student Involvement of iron handling proteins in bright light- induced stress in post hatch chick retina
Maurya Meenakshi, Nag Chandra Tapas, Kumar Pankaj, Roy Sankar Tara, Wadhwa Shashi
Phd-9 Anatomy Poorti Kathpalia
PhD Student Epigenetic regulation of Brain Derived Neurotrophic Factor in the auditory cortex of neonatal chicks exposed to prenatal rhythmic sound stimulation
Kathpalia Poorti, Nag Chandra Tapas, Sharma Arundhati, Roy Sankar Tara, Wadhwa Shashi
Phd-10 Anatomy Shivani Gupta PhD Student Attenuation of Acute Inflammatory Pain following a Surgical Incision by Neuropeptide Y in Rats
Gupta S, Gautam M, Kumar R, Prasoon P, Kaler S, Ray S.B.
Phd-11 Anatomy Binata Marik PhD Student Identification of Novel Mutations by Whole Exome Sequencing and Genotype-Phenotype Correlation in Patients with Refractory Rickets
Marik Binata, Bagga Arvind, Sinha Aditi, Hari Pankaj, Sharma Arundhati
Phd-12 Anatomy Kamakshi Mehta
PhD Student Attenuative role of Resveratrol on Arsenic trioxide induced cognitive deficits, cytoarchitectural alterations and oxidative stress in adult mice hippocampus.
Mehta K, Kaur B, Pandey KK, Dhar P
Phd-81 Anatomy Saba Sarwar PhD Student Ameliorative Effects Of Ascorbic Acid On Fluoride Induced Enteric Nervous System Developmental Defects
Sarwar S, Quadri JA, Singh S, Das P, Nag TC, Govind M, Roy TS, Shariff A
Phd-100 Anatomy Srishty Raman PhD Student Overexpression and purification of folded domain of cervical cancer related HPV-16 E7 protein
Srishty, Rawat Vandana, Yadav Chandra Subhash
Phd-108A
Anatomy Ekta Malik PhD Student Genetic screening of MEN1, AIP and GNAS1 genes in sporadic anterior pituitary adenoma cases
Malik Ekta, Jyotsna P Viveka and Sharma Arundhati
Phd-108B
Anatomy Ekta Malik PhD Student Significance of genetic testing for early diagnosis of Multiple Endocrine Neoplasia type-2 syndrome
Malik Ekta, Tandon Nikhil, Goswami Ravinder, Jyotsna P Viveka, Ganie Ashraf, Khadgawat Rajesh, Gupta Yashdeep and Sharma Arundhati
Phd-111 Anatomy Sushil Kumari PhD Student Descriptive picture on various aspects of acute Stevens-Johnson syndrome/Toxic epidermal necrolysis
Sangwan Kumari Sushil, Khanna Neena, Sharma Namrata, Agarwal Tushar, Sharma Arundhati
Phd-113 Anatomy Renu Singh PhD Student Association of the Genetic variations in monoamine pathway genes with Alcohol Dependence
Singh Renu, Gupta Ranjan, Grover Tripti, Ambekar Atul, Jain Raka, Sharma Arundhati
Phd-1
Title: AMELIORATIVE ROLE OF CURCUMIN ON BASAL FOREBRAIN OF MICE
SUBJECTED TO ARSENIC TRIOXIDE EXPOSURE
Name of the Authors: Kaur Balpreet, Mehta Kamakshi, Pandey Kumar Kamlesh and Dhar
Pushpa
Affiliation: Department of Anatomy
Presenting Author:
Name: Dr. Balpreet Kaur
Email: [email protected]
Corresponding Author:
Name: Dr. Pushpa Dhar
Email: [email protected]
Abstract
Introduction: Arsenic trioxide (As2O3) is administered as a chemotherapeutic agent for treating acute
promyelocytic leukemia. A number of adverse effects of As2O3 exposure on nervous system have been
reported, suggesting its role in targeting the nervous system. Basal forebrain (BF), with its cholinergic
neuronal population, is one of the various brain areas showing increased vulnerability towards a
number of insults. The present study was designed to determine the effects of As2O3 exposure on BF
of mice and to evaluate the effects of curcumin on As2O3 induced effects.
Aims & Objectives: To study the effects of curcumin supplementation on behavioral, biochemical &
immunocytochemical parameters pertaining to the basal forebrain of mice subjected to As2O3
exposure.
Material & Methods: Adult male mice were divided into control (I) and experimental such as As2O3
alone (II), Curcumin alone (III) and As2O3 and Curcumin cotreated (IV) groups. Groups II and IV
were further subdivided according to As2O3 dosage (2mg/kg bw; 4mg/kg bw). The test substances
were administered for 45 days. Morris Water Maze (MWM) test was performed on days 39-45 of the
experimental period. On day 46, perfusion fixed and freshly obtained brain tissue from control and
experimental animals was processed for immunohistochemical localization of various proteins and
estimation of oxidative stress markers respectively.
Result: In acquisition phase of MWM, As2O3 alone treated mice showed prolonged escape latency
and travelled longer distance to reach the platform whereas in probe trial, less number of platform
crossings were recorded. Significantly decreased glutathione levels and increased nitric oxide levels
were evident in As2O3 alone treated groups as against control and curcumin cotreated animals.
Immunoexpression of ChAT was less intense while that of GFAP and Iba-1 was more intense in
As2O3 alone treated animals compared to curcumin cotreated animals.
Conclusion: These preliminary observations are indicative of the beneficial role of curcumin
cotreatment on As2O3 induced effects on learning and memory, oxidative stress markers and
expression of various proteins with context to BF of mice.
Phd-2
MATURATION OF NEURONS AND ASTROGLIA IN DEVELOPING HUMAN COCHLEAR
NUCLEUS
Saini Shubhi
1, Jacob George Tony
1, Thakar Alok
2, Roy Kumar Kallol
3, Roy Sankar Tara
1
Department of Anatomy1, Department of Otorhinolaryngology
2, Department of Obstetrics
and Gynecology3, All India Institute of Medical Sciences, New Delhi-110029
Corresponding author- Tara Sankar Roy ([email protected])
Presenting author- Shubhi Saini ([email protected])
Introduction: It is not known when a fetus starts hearing in utero. There are reports of detectable
auditory brainstem responses at 18th week of gestation (GW). A mature auditory pathway is essential
for this. Cochlear nucleus (CN) is the first central relay station in the auditory pathway that is
responsible for transmitting and processing auditory information. In our previous study we have
shown that cochlear nerve myelinates around 20th GW. In the present study we examined the
maturation of the neurons and glia in the developing CN.
Aims & Objectives: To study the maturation and quantify the neurons and astroglia in cochlear
nucleus (CN) at different gestational and postnatal ages by using unbiased stereological methods and
expression of markers of neuronal (NeuN) and astroglial (GFAP) differentiation.
Materials and Methods: Brainstems from twenty-two aborted fetuses (10-32 weeks) and seven
postnatal infants were dissected, fixed, processed for cresyl violet staining and NeuN and GFAP
immunostaining. The total number of neurons and astroglia were estimated by the Optical
Fractionator probe on StereoInvestigator software (Microbrightfield Inc. VT, US). The data was
analyzed by dividing it into four groups (18-20 WG, 21-24 WG, 25-28 WG and >29 WG). Kruskal -
Wallis test followed by Dunn’s test for pair-wise comparison was used to compare the estimates.
Results: Qualitatively, Nissl granules in neurons and mature glial cells were observable from 20 WG.
The number of identifiable neurons (CV-stained and Neu-N-immunopositive) and astroglial cells
(GFAP-immunopositve) increased progressively from 18 WG. There was significant difference in
total neuronal counts between groups 1 and 3 (p=0.02) and 4 (p<0.0001) and between groups 2 and 4
(p=0.01). In NeuN-immunopositive neurons a statistical significance was observed between groups
1vs 3 (p=0.02) and 4 (p=0.001) and between groups 2 and 4 (p=0.02). In case of GFAP there was a
significant difference between groups 1 and 3 (p =0.04) and 4 (p= 0.001) and between groups 2 and 4
(p= 0.006).
Conclusions: In our study the qualitative observation and quantitative estimates of neurons and
astrocytes shows that there is a major growth phase of CN at 18 WG, which overlaps with the onset of
the physiological intrauterine responses to auditory stimulation. Therefore, this is a period of
vulnerability.
Phd-3
Yoga reduces disease activity and severity of comorbid depression and improves
quality of life in Rheumatoid arthritis: A randomized controlled trial
Gautam Surabhi, Kumar Uma, Kanga Uma, Dada Rima
Presenting Author:
Surabhi Gautam: Laboratory for Molecular Reproduction and Genetics, Department of
Anatomy, All India Institute of Medical Sciences, New Delhi, India
Email: [email protected]
Corresponding Author:
Rima Dada: Laboratory for Molecular Reproduction and Genetics, Department of Anatomy,
All India Institute of Medical Sciences, New Delhi, India
Email: [email protected]
Introduction:Besides routine medical therapy, recovery in Rheumatoid Arthritis (RA) is
dependent on several physical and psychological factors. Co-morbid depression adversely
affects RA outcome. Usual medical therapies with a limited scope fail to cure the
psychological component of the disease. So, mind body interventions have become the need
of hour in this age of super-specialization as majority of disease has a psychosomatic
component.
Aims & Objectives: This study aimed to assess impact of 8 weeks Yoga-based lifestyle
intervention (YBLI) on disease activity and depression severity in active RA patients on
routine disease modifying anti-rheumatic drugs (DMARDs) therapy.
Materials & Methods:A total of 72 RA patients were randomized into 2 groups: yoga (yoga
with DMARDs) and control (DMARDs only). Blood samples were collected pre and post
intervention for measurements of disease specific biomarkersfor systemic inflammation –
acute phase reactants (ESR, CRP), inflammatory cytokines (IL-6, IL-17A, TNF-α, TGF-β,
soluble HLA-G);neuroplasticity – BDNF, serotonin, β endorphins;cellular health –
oxidative stress (ROS, TAC), DNA damage (8OHdG), health span & longevity (SIRT1) and
cellular aging – telomerase activity and telomere length. Pre and post intervention of disease
activity score (DAS28ESR), functional status by health assessment questionnaire-disability
index (HAQ-DI), depression severity by Beck Depression Inventory II scale (BDI-II) and
quality of life(QoL) by World Health Organization Quality of Life (WHOQOL-BREF)
respectively were assessed.
Results:In YBLI group, there was reduction in mean levels of ESR (p<0.05*), CRP
(p<0.05***), ROS (p>0.05), TNF-α (p<0.05**), IL-6 (p<0.05**), IL-17A (p<0.05**) and
elevation in sHLA-G (p<0.05**) and TGF-β (p<0.05**) at 8 weeks compared to base line
level (day 0).YBLI participants showed significant improvements in DAS28 (p<0.05***),
HAQ (p<0.05***), VAS (p<0.05***) and WHOQOL-BREF scale (p<0.05***) in all 4
domains of physical health, psychological health, social relationships, and environmental
health over the control group.
Conclusion:Yoga practice results in regression of inflammatory processes by reducing
inflammatory cytokines, reducing pain perception, disability quotient, disease activity and
improved quality of lifein active RA patients. Thus yoga aids to settle relapses and prolong
the periods of remissionin RA. YBLI emphasizes immune-modulation may be beneficial for
individuals with RA as an adjunctive therapy.
Phd-4
IMPACT OF YOGA- AND MEDITATION-BASED LIFESTYLE
INTERVENTION ON DEPRESSION, QUALITY OF LIFE, AND CELLULAR
AGING IN INFERTILE COUPLES Madhuri Tolahunase#, Rajesh Sagarb, Rima Dada#
Affiliation: a Lab for Molecular Reproduction and Genetics,
Department of Anatomy, All India Institute of Medical Sciences, New Delhi,
India; b Department of Psychiatry, All India Institute of Medical Sciences,
New Delhi, India.
Presenting Author: Name: Tolahunase Rajaneesh Madhuri
Email: [email protected]
Corresponding Author:
Name: Dada Rima
Email: [email protected]
Introduction: Nearly half of the cases of infertility are of unknown origin comprising both
idiopathic and unexplained infertility. Depression, accelerated biological aging, and
decreased quality of life are associated with this group. Modern lifestyle plays a critical role
in both infertility and depression.
Aims & Objectives: The goal of this study was to evaluate the effect of yoga- and
meditation-based lifestyle intervention (YBLI) on depression, quality of life and cellular
aging in couples with infertility of unknown origin.
Design: A randomized controlled trial.
Materials and methods: Seventy-four couples diagnosed with infertility of unknown origin
were randomized into YBLI group (n = 37 couples) and routine expectant management
(REM) group (n = 37 couples). All couples were assessed both pre- and post-intervention
using Beck Depression Inventory-II scale (BDI-II), 26 item brief version of the World Health
Organization Quality-of-Life Scale (WHOQOL-BREF), and assay kits for the peripheral
blood biomarkers of cellular aging that included 8OH2dG, telomere length, telomerase
activity, ROS, TAC, Cox-II activity, cortisol, b-endorphin, DHEAS, IL-6, TNF-α, LL- 37,
sirtuin-1, BDNF, melatonin, and serotonin. Morphological and functional parameters of
semen were also evaluated in the male partners. WHO 2010 criteria were used for
morphological semen analysis. Functional assessment of the semen included cardinal markers
of cellular aging: Sperm DNA damage by DNA fragmentation index (DFI); oxidative stress
in semen by reactive oxygen species (ROS), and total antioxidant capacity (TAC); and
telomere metabolism in sperm by telomere length and telomerase activity).
Results:
Intention-to-treat analysis demonstrated that there was a significant reduction in depression
severity [difference between means, (95% CI)] in BDI-II score [-4.73 (-8.13, -3.47), p < 0.
001] after YBLI compared to REM group. The study also showed significantly decreased
cellular aging and improved quality of life (in all four do- mains of WHOQOL-BREF scale -
physical, psychological, social, and environmental) in YBLI group in comparison to REM
group (all p < 0.05). Semen analysis in male partners showed a significant decrease in DNA
damage and a significant optimization of oxidative stress in YBLI group compared to REM
group (all p < 0.01).
Conclusion:
The results showed that YBLI has an important role in decreasing depression,
delaying/reversing cellular aging, and increasing quality of life in couples with infertility of
unknown origin. In addition, improvement in the morphological and functional semen
parameters of male partners in YBLI group suggest that cellular aging and depression are
associated with infertility, and yoga and meditation may increase chances of natural
conception and healthy next generation by improving gamete structure and function.
Phd-5
Title: Impact of yoga based lifestyle intervention on sperm oxidative DNA damage: study on
fathers of non-familial sporadic heritable retinoblastoma patients
Shilpa Bisht1, Bhavna Chawla
2, Rima Dada
1
Affiliation: 1Laboratory for Molecular Reproduction and Genetics, Department of
Anatomy, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
2Ocular Oncology and Paediatric Ophthalmology Service, Dr.Rajendra Prasad Centre for
Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi,
India.
Presenting Author:ShilpaBisht
Email:[email protected]
Corresponding Author: Professor Rima Dada
Email:[email protected]
Abstract Body
Introduction:Sperm chromatin integrity is essential for fertilization, proper embryonic
development and birth of healthy offspring. Sperm is highly vulnerable to oxidative damage
to both nuclear and mitochondrial DNA due to minimal cytosolic anti-
oxidants.Retinoblastoma is the most common childhood malignancy where the de novo germ
line mutations are mainly inherited from paternal allele.
Aims & Objectives: The present study is planned as a case control study on the fathers of
children affected with non-familial sporadic heritable retinoblastoma to see the effect of yoga
based lifestyle intervention on the decline in oxidative stress and oxidative DNA damage.The
present study was planned as a case control study with 75 fathers of children affected with
non-familial sporadic heritable retinoblastoma cases and 50 fathers of healthy children were
enrolled as healthy controls. Study duration was 6 months.
Materials and Methods:Fathers of children affected with non-familial sporadic heritable
retinoblastoma were enrolled in a yoga-based lifestyle intervention programme. Semen
samples were collected at base line (day 0), after 1 month, 3 months and 6 months of yoga
intervention. ROS, DNA fragmentation index (DFI) and 8-hydroxy-2’-deoxyguanosine (8-
OHdG) levels were estimated at base line (day 0), 1 month, 3 months and 6 months duration.
ROS, DFI, 8-OHdG were also assessed in fathers of healthy children.
Results: Semen parameters were assessed as per the WHO, 2010 guidelines. The seminal
mean ROS levels (p<0.05), sperm DFI (p<0.001), 8-OHdG (p<0.01) levels were significantly
higher in fathers of children withnon-familial sporadic heritable retinoblastoma as compared
to fathers of healthy children. There was a reduction in mean DFI levels post yoga-based
lifestyle intervention at 1 month (p=0.63), 3 months (p=0.046) and 6 months (p=0.032).
Similarly, there was a reduction in seminal mean ROS levels at 1 month (p=0.74), 3 months
(p=0.039) and 6 months (p=0.006). The levels of DNA oxidative base adduct i.e., 8-OHdG
were also significantly reduced at 1 month (p=0.68), 3 months (p=0.043) and 6 months
(p=0.022) with respect to its base line levels (day 0).
Conclusion: Yoga-based simple lifestyle intervention meditation may significantly lower
oxidative stress and oxidative DNA damage, and levels of mutagenic base 8-OHdG in the
sperm DNA. Thus, yoga-based intervention is alternative and complementary treatment for
maintaining/restoring sperm DNA integrity and hence, can reduce the prevalence of
childhood cancers where poor sperm DNA integrity could be one of the reason.
Phd-6
TITLE: SPERM TRANSCRIPT SIGNATURE IN EARLY PREGNANCY LOSS
Authors: Vidhu Dhawan, Neena Malhotra, Nita Singh, Vatsala Dhadhwal, Rima Dada
Presenting author- Vidhu Dhawan
Email: [email protected]
Corresponding author- Rima Dada
Email [email protected]
Introduction: Paternal factors have been cited as an important contributor in the normal
embryonic development. Derangements in the sperm genomic integrity, oxidative stress and
dysregulation in gene expression have been associated as a causal factor for esrly pregnancy
loss in both spontaneous and assisted conceptions.
Objectives: The present case control study was designed to evaluate the expression of genes
critical for embryonic development. The evaluation of seminal oxidative stress, DNA damage
and 8-OHdG was also done.
Methods: Semen sample was obtained from male partners of couples experiencing recurrent
pregnancy loss (RPL) (n=75), and recurrent implantation failure (RIF) (n=75) patients and
healthy fertile controls (n=75). q-PCR analysis was done to assess the expression of SOX3,
STAT4, RPS6, RBM9, RPL10A, FOXG1, TOMM7, EIF5A as well as OGG1 and PARP1 after
normalization withGAPDH and β-actin. ROS levels (RLU/sec/million sperm), DNA
fragmentation index (%) and 8-OHdG levels were estimated.
Results: The relative expression of RPS6 and RBM9 was found to differ significantly
between RPL patients and controls, while expression of RPS6, RBM9, RPL10A and TOMM7
was seen to differ significantly between RIF patients and controls. SOX3 and PARP1 showed
downregulation in RPL patients, while all the other genes in RPL and RIF patients showed
upregulation. The median ROS level was seen to be higher (>29) in RPL [40.9 (3.1-731.24)]
and RIF patients [57.75 (10.06-1186.9)] w.r.t controls [17.891.15-53.90] (p<0.001). The
mean DFI levels were seen to be higher (>31) in RPL (34.08 ± 5.27) and RIF patients (36.14
± 5.01) w.r.t controls (27.8 ± 4.02) (p<0.001). 8-OHdG levels were found to be higher in both
RPL and RIF patients (p<0.001).
Conclusion: Derangements in sperm RNA expression and genomic integrity pose as a
critical determinant of embryo viability and affect the pregnancy outcomes and even affect
future health of the progeny.
Phd-7
Age-related human spiral ganglion neuronal loss assessed by unbiased stereology
Kaur Charanjeet1, Jacob Tony George
1, Nag Tapas Chandra
1, Thakar Alok
2, Bhardwaj
Dayanand3, Roy Tara Sankar
1
1Departments of Anatomy,
2Otorhinolaryngology and
3Forensic Medicine and Toxicology,
All India Institute of Medical Sciences, New Delhi, India
Presenting Author: Charanjeet Kaur ([email protected])
Corresponding Author: Tara Sankar Roy ([email protected])
Introduction: Age-related hearing loss (presbycusis) is the most prevalent neurodegenerative
condition in our aged population. Literature suggests that oxidative stress, mitochondrial
mutations, excitotoxicity caused by neurotransmitters or may be other environmental factors,
ultimately results in loss of SGNs with aging. Therefore, estimation of total number of
neurons in the SG at various ages is important as baseline information. Neurons are easily
identified in Nissl-stained (cresyl violet) sections or by the expression of specific neuronal
markers like the calcium-binding protein Parvalbumin (PV).
Aim: To estimate and compare number of human SGNs identified by CV staining and PV-
expression at various ages using unbiased stereological methods.
Materials and methods: Thirty-five human temporal bones were obtained from individuals
who had died of various causes, without any known history of inner ear disease or hearing
loss, from the mortuary after obtaining clearance from the Institutional Ethics Committee.
These bones were fixed in 4% paraformaldehyde (0.1M PB, pH 7.4), decalcified in EDTA,
cryoprotected and sectioned to obtain 30 µm thick serial coronal sections. Every 7th
section
was stained with cresyl violet (CV, 1%) and immunostained for the expression of
parvalbumin (ab11427, 1:5000) using standardized protocol. CV-stained and PV-
immunostained sections were used for estimating the total number of neurons with the optical
fractionator probe using StereoInvestigator software (Microbrightfield Inc. VT, USA). The
relationship between age and total number of SGNs was made with linear regression analysis.
Results: The proportion of SGNs (either stained by CV or PV) were significantly and
negatively correlated with increasing age (slope -283.3 F (1, 33) = 67.63, p< 0.0001 with an
r2 of 0.67) and (-263.9 F (1, 33) = 47.28, p < 0.0001) in case of CV-stained and PV-ir SGNs,
respectively. Further, there was no significant difference between the two staining methods
on applying Bland-Altman plot.
Conclusion: The present study demonstrates that the total number of the human SGNs (Nissl
stained) and SGNs immunoreactive for PV decreases with ageing and our results demonstrate
that this process begins early.
Phd-8
Involvement of iron handling proteins in bright light- induced stress in post
hatch chick retina
Maurya Meenakshi *, Nag Chandra Tapas, Kumar Pankaj, Roy Sankar Tara, Wadhwa Shashi
Department of Anatomy, All India Institute of Medical Sciences, New Delhi 110029, India
Presenting author: Meenakshi Maurya
E mail: [email protected]
Corresponding author: Dr Tapas Chandra Nag
E mail: tapas_nag @aiims.edu
Introduction: The importance of iron in nutrition and cellular metabolism has been
recognized for over a century. However, excess of cellular iron causes generation of reactive
oxygen species (ROS) that play a detrimental role in the pathophysiology of various diseases.
It is known that dysregulation of iron regulatory proteins (IRP) leads to such altered iron
homeostasis, leading to pathophysiology. On the other hand, a neuroprotective role of IRP
has emerged from many studies. For example, exposure to intense light intensities that causes
damage to the retina is coupled with increased expression of ceruloplasmin (Cp) in retinal
glia. Iron transporters, such as transferrin (Tf), transferrin receptor-1 (TfR-1) and Cp are
involved in iron regulation. However, the role of iron transporters in light-induced retinal
damage is unknown.
Aim and objective: The aim of this study was to understand if bright light causes oxidative
stress and if so, how iron transporters are involved in bright light-induced damage.
Materials and Methods: One day-old chicks were reared in normal 12 h light -12 h dark
cycle (12L: 12D) for 7 days (400 lux). From day 8 (0h) onwards, they were exposed to high
intensity light (5000 lux, experimental) and 400 lux (control). At 168h of exposure, retinal
samples were collected. Lipid peroxidation levels in retinal extracts were estimated by
Thiobarbituric acid reactive substances (TBARS) assay. Cryosections were immunolabeled
for Tf, TfR-1 and Cp and their levels determined by immunoblotting. Ultrastructural changes
were examined by transmission electron microscopy (TEM).
Results: After exposure of 5000 lux with respect to 400 lux light intensity at 168h time point
under 12L: 12D photoperiod, TBARS assay revealed an increase in lipid peroxidation level in
5000 lux vs 400 lux exposed group. Immunohistochemical localization of Tf was intense in
photoreceptors, inner nuclear layer (INL), ganglion cell layer (GCL) and Müller cell
processes. TFR-1 was extensively expressed in outer plexiform layer, INL and inner
plexiform layer (IPL). Western blot analysis revealed increased expression of Tf (p=0.03) and
TfR-1(p=0.04), whereas Cp expression was decreased (p=0.03) in 5000 lux vs 400 lux
exposed groups. TEM revealed degenerated outer segments and disorganized mitochondrial
cristae in photoreceptor cells. There were organelle depletion in INL cells, gliosis of Müller
cell processes and vacuolated axons in IPL.
Conclusion: Increased expressions of Tf and TfR-1 after bright light exposure implicate that
retinal iron regulation is perturbed by bright light, while a downregulation of cp is supposedly
associated with increases in free iron levels that generate ROS, as reflected in increased lipid
peroxidation levels. Presence of necrotic cells in INL indicates the adverse, secondary effect
of light after photoreceptor damage. Such alterations implicate the pivotal role of IRP in the
protection of the retina from light-induced degeneration, most likely mediated via altered
retinal iron levels.
Phd-9
Abstract for AIIMS Research Day 2019
Epigenetic regulation of Brain Derived Neurotrophic Factor in the auditory cortex of
neonatal chicks exposed to prenatal rhythmic sound stimulation
Kathpalia Poorti, Nag Chandra Tapas, Sharma Arundhati, Roy Sankar Tara, Wadhwa
Shashi
Department of Anatomy, All India Institute of Medical Sciences, New Delhi
Presenting author: Poorti Kathpalia, [email protected]
Corresponding author: Prof Tapas Chandra Nag, [email protected]
Introduction: The beneficial effect of rhythmic sound stimulation on the developing brain of
fetuses is known for quite a long time. Experimental studies have shown that prenatal music
stimulation increases the neuronal number in the hippocampus and auditory cortex in
neonatal chicks. Music also increases the levels of brain derived neurotrophic factor (BDNF)
and its receptor (TrkB) in the hippocampus of animals when the intensity of sound is
moderate (60 dB), which might be an underlying factor for increased neuronal number
detected in neonatal brain. However, the effect of high intensity prenatal sound (noise) on the
growing brain, as is commonly experienced by many pregnant women in modern societies,
has not been studied with respect to the expression and regulation of BDNF, a key molecule
involved in early neurogenesis.
Aims and Objectives: We examined the effect of high intensity (110 dB) prenatal rhythmic
sound stimulation on BDNF levels and its regulation in neonatal auditory cortex via
immunohistochemical localization of BDNF and quantification of the expressions of BDNF
protein and mRNA levels, acetylated histone H2B and H4 levels at the BDNF promoter
region in neonatal chick auditory cortex exposed to prenatal high intensity sound.
Materials and Methods: Fertilized eggs of white leghorn chickens were exposed to
rhythmic sound (sitar music, 100-4000 Hz)/noise (traffic noise, 30-3000 Hz with a peak at
2700 Hz) at 110 dB from embryonic day 10 (E10) until hatching. The control eggs did not
receive any sound stimulation. Thus, the study had three groups (n=21/group): Control,
Music, Noise. Post-hatch neonatal chick brains were dissected and the auditory cortex
chopped out. Immunohistochemistry (IHC), immunoblotting, real time PCR and chromatin
immunoprecipitation were performed from the brain samples by standard methods.
Results: BDNF immunopositive neurons were observed in neonatal auditory cortex by IHC.
An increase in proBDNF levels was found in prenatal noise exposed auditory cortex with
respect to music exposed chicks. A decrease in BDNF transcript was seen in the auditory
cortex after prenatal sound stimulation, irrespective of the nature of the sound. The levels of
acetylated histone H4, but not that of H2B, were increased at the BDNF promoter region.
Conclusion: BDNF expression in the neonatal auditory cortex was found to be epigenetically
regulated in response to prenatal rhythmic sound stimulation. An enrichment of acetylated
Histone H4 was found at BDNF promoter region in the neonatal auditory cortex. The
biological significance of this remains to be determined in future studies (supported by grants
from CSIR, New Delhi (No. 37(1420)/10/EMR-II).
Phd-10
Title: Attenuation of Acute Inflammatory Pain following a Surgical Incision by
Neuropeptide Y in Rats
Author: Gupta S, Gautam M, Kumar R, Prasoon P, Kaler S, Ray S.B.
Affiliation:
Presenting Author: Shivani Gupta, email id: [email protected]
Corresponding Author: Subrata Basu Ray, email id: [email protected]
Department of Anatomy, All India Institute of Medical Sciences, New Delhi – 29, India
Aim: Postoperative pain continues to be an important health care problem despite substantial
advancement in understanding the mechanisms responsible for initiation and transmission of
pain. A-delta and C nerve fibers transmit the pain signals to the spinal cord, where these are
modulated by various neurotransmitters and neuropeptides. Neuropeptide Y (NPY) is
abundantly distributed in the mammalian nervous system. Several reports have shown its
involvement in acute and chronic preclinical pain models but its role in postoperative pain is
poorly understood. In the current study on the hind paw incision rodent model, the following
were investigated – temporospatial changes in expression of NPY in the spinal cord,
antinociception if any, after intraspinal administration of synthetic NPY and its reversal by
NPY antibody.
Methods: Male Sprague-Dawley rats were subjected to hind paw incision. The spinal cords
were collected (n=48) after euthanasia and processed for immunohistochemical labelling of
NPY using the avidin-biotin complex method. Another set of animals (n=12) were
administered NPY or saline through an intrathecal catheter. Finally, NPY antibody (n=6) was
administered through catheter followed by NPY and the effect observed. Three nociceptive
assays (Guarding behavior, mechanical allodynia and thermal hyperalgesia) were used to
evaluate the antinociceptive effect starting from 2h post-incision until postoperative day 7.
Results: NPY immunoreactivity was observed as punctate varicosities in the superficial
laminae of the dorsal horn. On the contrary, neurons positive for NPY, was observed in the
deeper laminae. NPY immunostaining decreased after incision at 3 h followed by an increase
at 12 h. At day 1, it decreased again. This variable pattern of expression suggested the
involvement of NPY in postincisional nociception. Subsequently, on intrathecal
administration, nociception was significantly decreased between 2 h to day 2, which was
reversed by antibody to NPY.
Conclusion: Neuropeptide Y likely acts as an antinociceptive factor in the spinal modulation
of pain. This information could have clinical relevance.
Phd-11
Identification of Novel Mutations by Whole Exome Sequencing and Genotype-
Phenotype Correlation in Patients with Refractory Rickets
Marik Binata
1, Bagga Arvind
2, Sinha Aditi
2, Hari Pankaj
2, Sharma Arundhati
1*
1Departments of Anatomy and
2Pediatrics, AIIMS, New Delhi
Presenting author: Binata Marik, PhD Student, [email protected],
*Corresponding author: Prof. Arundhati Sharma, [email protected],
Introduction: Vitamin-D deficiency rickets is common and treatable with vitamin D.
Refractory Rickets (RR), a genetic disorder resistant to vitamin D, includes entities like
Hypophosphatemic Rickets (HR), Renal Tubular Acidosis (RTA), Vitamin D Dependent
Rickets (VDDR), Skeletal Dysplasias, Polycystic Kidney Disease (PKD), malabsorption and
Chronic Kidney Disease (CKD). Diagnosis and management of RR is extremely difficult due
to the similarity of clinical features and different treatment modalities. Genetic screening is
crucial for a definitive diagnosis. Genetic confirmation at an early age leads to timely
initiation of treatment that improves growth potential and reduces skeletal deformities.
Aim: To identify mutations and perform genotype-phenotype correlation in patients with RR.
Materials and methods: A total of 46 patients belonging to 36 families were recruited.
Detailed clinical features and family history were documented, and five milliliters blood
samples collected after taking informed consent. Whole Exome Sequencing (WES) was
performed for the patients and 4 controls, and data analyzed using standard bioinformatics
approach. Mutations identified were validated by Sanger sequencing in 100 controls.
Results: Of 46 patients, 24 were males and 22 females with one family reporting
consanguinity. Patients had short stature, skeletal deformities, and hypophosphatemia with
mean age of onset at 2.8+1.3 years. WES revealed 38 different mutations in 12 genes (PHEX,
DMP1, CLCN5, ATP6V1B1, SLC4A1, SLC2A2, CYP27B1, VDR, SLC34A1, PKD1, SLC52A1
and COL11A2) in the 46 patients of which 17 mutations were novel.
Genetic testing helped in delineating the cases as HR (PHEX-41.30%, DMP1-4.34%), Dent
disease (CLCN5-2.17%), Distal RTA (SLC4A1-8.69%, ATP6V1B1-8.69%), Fanconi-Bickel
syndrome (SLC2A2-6.52%), VDDR type-1A (CYP27B1-8.69%), VDDR type-2A (VDR-
10.86%), Hypophosphatemic nephrolithiasis/Osteoporosis type 1 (SLC34A1-2.17%), PKD
(PKD1-2.17%), Riboflavin deficiency (SLC52A1-2.17%) and Otospondylomegaepiphyseal
dysplasia (COL11A2-2.17%) based on the mutations identified. Variability in disease severity
was observed wherein patients having missense PHEX mutations (c.1601C>Tp.P534L,
c.1970A>Gp.Y657C) were moderately affected with delayed onset of disease and healing
rickets, while patients carrying PHEX deletions/insertions
(c.566_567delAGp.Q189Hfs*22, c.651_654delACATp.H128fs*2,
c.1336_1337insAATA p.F446*) showed a severe phenotype with growth retardation in
addition to undergoing osteotomy to correct limb deformities. Deletions/insertions caused
frameshift resulting in premature termination leading to truncated/non-functional PHEX
protein. Intrafamilial variability in disease severity was also observed, in one family where
the female patient had short stature, limb deformities and delayed teeth eruption but her
mother presented only with short stature. Genetically, this variability is due to the presence of
two PHEX mutations in the patient but a single PHEX mutation in the mother. In another
family where the proband and his brother were born of consanguineous marriage,
homozygous VDR mutation (c.298C>Tp.R50*) was seen while their unaffected parents
were heterozygous carriers. In a family with suspected Fanconi syndrome or Dent disease,
compound heterozygous SLC2A2 mutations were identified in the patient who showed
phosphaturia, glycosuria, hypercalciuria and beta-2-microglobulinuria suggesting Fanconi-
Bickel syndrome. Sanger sequencing validation revealed that the mutations segregated in the
family; one from each parent.
Conclusion:
Genetic testing identified different types of RR confirming diagnosis, allowing treatment
initiation and timely management. Studies on more patients will help in designing a targeted
gene panel for rapid screening and accurate diagnosis of RR.
Phd-12
Title: Attenuative role of Resveratrol on Arsenic trioxide induced cognitive deficits,
cytoarchitectural alterations and oxidative stress in adult mice hippocampus.
Introduction: Exposure to arsenic (As) through consumption of contaminated drinking water
is associated with affliction of various organ systems. The reports of cognitive deficits in both
humans and in rodent models following exposure to iAs are suggestive of targeted influence
of As on CNS. The beneficial effects of naturally occurring antioxidants have been explored
in various neurodegenerative disorders, metal toxicities, and oxidative stress injuries.
Resveratrol with its ability to cross the blood–brain barrier, is one such antioxidant.
Aims and Objectives: The present study evaluated the effects of Resveratrol on cognitive
functions, biochemical parameters and cytoarchitectural alterations with context to mice
hippocampus (CA-1) following As2O3 administration.
Materials and Methods: Adult female mice of reproductive maturity (monitored by estrus
cycles) were divided into control and experimental groups. As2O3 (2 and 4 mg/kg bw) alone
or along with Resveratrol (40 mg/kg bw) was administered orally to experimental groups for
45 days whereas the controls received either no treatment or vehicle for Resveratrol (Gum
acacia). From Day 33 to 46 of the experimental period, a battery of behavioural test (OFT,
EPM and MWM) was carried out. On Day 46, the animals were sacrificed either by perfusion
fixation or by euthanasia and the brain tissues, thus obtained were processed for observing
cytoarchitectural features and analyzing GSH (Reduced glutathione) and NO (Nitric oxide)
levels respectively.
Results: Behavioural tests revealed enhanced anxiety levels and impairment of cognitive
functions (learning and memory) in As2O3 alone exposed groups. Oxidative stress parameters
(GSH and NO) in hippocampal tissue of As2O3 (2mg/kgbw) alone exposed animals were
altered. Significant reduction in layer (Stratum Pyramidale) thickness along with decrease in
neuronal density and neuronal area of Pyramidal cells in As2O3 alone exposed animals was
observed as compared to controls and Resveratrol supplemented animals.
Conclusion: These observations provide preliminary evidence of protective role of
Resveratrol on functional, biochemical and structural parameters of hippocampus in mice
subjected to chronic As2O3 exposure.
Phd-81
Ameliorative Effects Of Ascorbic Acid On Fluoride Induced Enteric Nervous System Developmental Defects
Sarwar S1, Quadri J A1, Singh S1, Das P2, Nag T C1, Govind M3, Roy T S 1, Shariff A1
1Department of Anatomy, 2Department of Pathology,3 Department of Gastroenterology
All India Institute of Medical Sciences, New Delhi, India
INTRODUCTION: In fluorosis endemic areas gastrointestinal complications including
constipation, infrequent diarrhea, loss of appetite and abdominal pain which mimic the
Irritable Bowel Syndrome (IBS) are very common. Severe anemia in fluoride exposed
pregnant women is very frequent and considered to be manifestation of compromised
intestinal absorption due to fluoride induced gut mucosal injury.[1] Enteric Nervous System
(ENS), plays a central role in gut functions including peristalsis, digestion, nutrient
absorption, stomach & rectal emptying. Any ENS abnormality may lead to Patho-
physiological conditions. Fluoride is known as a induced oxidative stress. Due to oxidative
stress Enteric neurons may altered the ENS. Vitamin C decreased the oxidative stress it is
known so it is hypothesized that ascorbic acid prevent the ENS due to fluoride toxicity. ENS
contains various types of neuronal and non-neuronal (glial) cells. These cells include enteric
glia, nitrergic and cholinergic neurons[2.
AIM-The aim of the study was to evaluate the ascorbic acid effects on fluoride induced
changes in hind-gut and its impact on ENS.
METHODOLOGY-Female and Male wistar rats were taken and allowed to mate.Divided into
three groups, Control,100ppm fluoride and 100ppm+asorbic acid. During the pregnancy and
lactation (up to post natal 20th days) administered with 100 ppm of fluoride and
100ppm+ascorbic acid. After birth pups were allowed to be on mother milk for 20th-days
and kept in the same cage, allowed to drink same drinking water. The pups were euthanized
on 20th post-natal days and gut tissues were collected for different analysis, such as
histopathology ,real time Pcr, oxidative stress , immunofluorescence ,and ultrastructure.
RESULTS: In fluoride administered groups shows histopathological changes in gut tissue and
enteric ganglion.
Immunofluorescence result showed that changes in the number of HUCD, nNOS neurons
and Glial cells.
Real time PCR result showed that the expression of mRNA encoding HUCD, nNOS, and GFAP
receptors decreased in fluoride treated groups but in ascorbic acid with fluoride treated
groups shows that expression of mRNA encoding HUCD nNOS and GFAP receptors were
increased in comparison to fluoride treated groups .Level of LPO and SOD is increased in
fluoride treated group in comparison to control. but in ascorbic acid treated group it was
decreased in comparison to fluoride treated groups. ENS cells (enteric neurons and glial
cells) showed various cellular and sub-cellular ultrastructural changes. These fluoride
induced ultrastructural alterations include heterochromatic nucleus, ENS cells showed
dilated RER, ER fragmentation, mitochondrial disintegration and nuclear membrane
fragmentation but in ascorbic acid treated group shows preventive effects.
CONCLUSION: Fluoride induces adverse effects on ENS by neuronal cell injury and apoptosis
and ascorbic acid shows protective effect.
Presenting Author: Saba Sarwar, PhD student, Department of Anatomy AIIMS New Delhi,
Corresponding -Author: Professor A Shariff, Department of Anatomy, AIIMS, New Delhi,
Phd-100
Title:Overexpression and purification of folded domain of cervical cancer related HPV-16 E7
protein
Name of authors:Raman Srishty; RawatVandana; YadavChandra Subhash
Affliation: PhD student, AIIMS, New Delhi
Presenting author:Srishty Raman, PhD student, Department of Anatomy, AIIMS, New Delhi
Email:[email protected]
Corresponding author: Dr Subhash Chandra Yadav, Assistant Professor, Department of Anatomy,
AIIMS, New Delhi
Email:[email protected]
Abstract body
Introduction: Overexpression of antigenic epitopes/ domains of a potent biomarker protein using
recombinant DNA technology for generation of antibodies against them, can be a boon to the
diagnostic industry. However, the idea brings along a great challenge with itself because individual
epitopes tend to accumulate as insoluble aggregates when expressed separately. Besides, identifying
the right sequence for a domain to be able to fold independently, the choice of an expression vector
plays an important role in understanding the folding pattern and underlying protein-engineering
events to isolate the functional domains of a protein.
Aims & Objectives: To obtain individual domain proteins of HPV-16 E7 protein in purified folded
form and related insights on protein engineering events, which may be used to generate such
domain specific polyclonal antibodies for enhancing the sensitivity of ELISA.
Materials & methods: In silico analysis for identification of individual domain sequences was
performed using various boinformatic tools. Each individual domain along with the full-length E7
protein of HPV-16 were overexpressed in different bacterial vector systems- pET21a, pET28a, pET
sumo and purified to obtain their folded forms using affinity chromatography. The proteins were
further confirmed by western blotting experiment using domain specific antibodies. Biophysical
characterization using tryptophan fluorescence and circular dichroism was done to confirm folding
pattern.
Results:Full-length E7 protein was overexpressed in soluble form in pET21a while both the domains accumulated as insoluble aggregates in pET21a and pET28a. However, the domains did express well in pET sumo.The content of secondary structure predicted by insilico analysis was similar to the results obtained by far UV CD. Conclusion:It was found that the greater the tag, the lesser were the chances of the epitopes to
accumulate as insoluble aggregates, and hence SUMO-tagged domains were obtained in a greater
fraction in soluble form as compared to HIS-tagged domains. This study may further help producing
similar folded domain of many other proteins which can be used to increase the sensitivity of various
antibody based diagnostic procedures.
Phd-108A
Title: Genetic screening of MEN1, AIP and GNAS1 genes in sporadic anterior pituitary adenoma cases Malik Ekta1, Jyotsna P Viveka2 and Sharma Arundhati1
Affiliation: lLaboratory of Cyto-molecular Genetics, Department of Anatomy, AIIMS, New Delhi, India 2Department of Endocrinology and Metabolism, AIIMS, New Delhi, India Presenting author: Name: Ekta Malik Email: [email protected] Corresponding author: Name: Dr. Arundhati Sharma Email: [email protected] Introduction: Anterior pituitary adenomas (APA) being benign intracranial neoplasms of the adenohypophysis are usually sporadic. They are categorized as micro (<10 mm) and macroadenoma (>10mm) based on size. Hormonal dysfunction is the hallmark of APA which are classified into functioning (Cushing disease, acromegaly, prolactinoma) and nonfunctioning adenoma. Various tumor suppressor genes like MEN1, AIP and GNAS1 can be associated with these adenomas. Aim: To screen MEN1, AIP and GNAS1 genes for mutations in sporadic APA. Materials and Methods: 100 clinically diagnosed APA patients from the outpatient department of Endocrinology and Metabolism and 50 healthy individuals from the general population were recruited. Detailed clinical and family history was noted and five ml blood collected after taking informed consent. Genomic DNA was subjected to PCR amplification of all the exons of MEN1, AIP and GNAS1 genes, followed by direct sequencing. Results were compared with reference sequences in the public databases and prediction of functional effects of the variations was done using bioinformatics tools Mutation t@ster and Human Splicing Finder (HSF). Statistical analysis was done by SPSS v21.0 software. Results: Out of 100 patients (75 functioning and 25 nonfunctioning), 56 were females and 44 males. Of the adenomas, 65 were macroadenoma and rest microadenoma and most (n=87) of the tumors were
nonaggressive. Mean age and age at onset of APA patients was 34.00±12.44 and 25.04±12.04 years respectively. Genetic screening revealed 3 intronic and 1 exonic variations in MEN1 and 2 intronic changes in AIP gene. MEN1 changes identified were synonymous variation L404L (1 patient), reported intronic variations c.913-79T>A (2 patients), c.784-129T>A (1 patient) and c.913-42G>C (2 patients) and SNP rs669976 in 41 patients. SNP rs669976 was found to be significantly associated with size of adenoma (p-0.042) which showed that patients with SNP rs669976 had more macroadenomas as compared to other SNPs. SNPs rs654440 (60% cases; 70% controls) and rs2071313 (70% cases; 70% controls) were also found in MEN1 gene. SNP rs654440 showed borderline significant association with age at onset (27.73±11.79) (p-0.078) which revealed that the patients who were having SNP rs654440 had lower age at onset as compared to patients with the normal allele. AIP screening revealed reported intronic splice site variations c.100-6C>A and c.100-18C>T in 1 patient each, SNPs rs2276020 (40% cases, 25% controls), rs641081 (55% cases, 10% controls) and rs4084113 (68% cases, 18% controls). Variations c.913-42G>C (MEN1) and c.100-6C>A (AIP) were predicted to be damaging by mutation t@ster and MEN1 variations c.913-79T>A, c.784-129T>A, c.913-42G>C and variation c.100-6C>A of AIP were found to alter the splice site as predicted by HSF. While no SNPs of AIP showed significant association with any clinical and histopathological features, no variations were found in the GNAS1 gene. Conclusion: MEN1 and AIP variations in sporadic APA were mostly intronic or at intron-exon junctions and seem to alter splicing. Preliminary analysis suggests that splicing affects the final
transcript and results in altered protein thereby leading to the mechanism of tumor
causation.
Phd-108B
Title: Significance of genetic testing for early diagnosis of Multiple Endocrine Neoplasia type-2 syndrome Malik Ekta1, Tandon Nikhil2, Goswami Ravinder2, Jyotsna P Viveka2, Ganie Ashraf2, Khadgawat Rajesh2, Gupta Yashdeep2 and Sharma Arundhati1
Affiliation: lLaboratory of Cyto-molecular Genetics, Department of Anatomy, AIIMS, New Delhi, India 2Department of Endocrinology and Metabolism, AIIMS, New Delhi, India Presenting author: Name: Ekta Malik Email: [email protected] Corresponding author: Name: Dr. Arundhati Sharma Email: [email protected] Introduction: Multiple Endocrine Neoplasia-2(MEN2) is a rare and complex genetic syndrome inherited as an autosomal dominant trait. It is characterized by co-occurrence of tumors in multiple endocrine glands like thyroid (Medullary Thyroid Carcinoma), adrenal (pheochromocytoma) and parathyroid (hyperparathyroidism) glands. The di erent combination of the endocrine tumors with or without nonendocrine tumors gives rise to di erent variants of MEN2: MEN 2A, MEN2B and Familial MTC (FMTC). MEN2A is the most common form (70-80%) which includes MTC, pheochromocytoma and HPTH. MEN2B is the least common (5%) characterized by association of aggressive MTC, pheochromocytoma, Marfanoid habitus, mucosal and intestinal ganglioneuromatosis. FMTC constitutes approximately 10-20% cases of MEN2 and MTC is the only clinical manifestation. RET, a protooncogene is the candidate gene associated with MEN2. It has 21 exons and encodes a tyrosine kinase transmembrane receptor, located on chromosome 10q11.2. RET protein is involved in cell signaling and triggers chemical reactions that instruct cells to respond to their environment by dividing. Mostly, mutations occur in two regions of the RET protein - the cysteine-rich extracellular domain and the tyrosine kinase domain. Consequences of these mutations depend on the region of the RET gene harboring mutations.
Aim and Objective: The present study aimed to identify RET mutations in clinically suspected patients with MEN2 syndrome. Materials and methodology: A total of 65 patients of MEN2 were recruited from the department of “Endocrinology and Metabolism”, AIIMS for genetic screening. Detailed clinical and family history was noted and 5ml peripheral blood samples collected for DNA extraction after taking informed consent. The extracted DNA was subjected to PCR amplification of RET gene followed by direct sequencing and results compared with reference gene sequences in the public databases. Results: Out of the 65 patients screened, 37 were females and 28 males with mean age 39.67±12, mean age at onset 32.88±11.78 years and 15 had a positive family history. All the 65 patients had undergone thyroidectomy. Hotspot mutations were found in 33 patients. Exon 11 mutations C634R and C634Y were found in 22 and six patients respectively. Exon 10 mutation C618S and exon 16 change M918T was found in two patients each while C618R (exon 10) in one patient. Patients with MEN2A and FMTC had mutations (C634R, C634Y, C618Sand C618R) in the cysteine rich region while MEN2B patients had mutation M918T (exon 16) in the tyrosine kinase domain. The results are in accordance with the North American Neuro Endocrine Tumor Society consensus guidelines wherein risk of aggressiveness and metastasis is high in MEN2A and FMTC patients with mutations in cystein rich regions while MEN2B with codon 918 mutation are the most aggressive of all with metastasis in the first few years of life. Conclusion: Genetic testing is crucial for confirming the diagnosis and determining the type of MEN2. It is also important for early diagnosis, timely treatment/management and carrier identification in family
members for detection of at risk individuals.
Phd-111
Title: Descriptive picture on various aspects of acute Stevens-Johnson
syndrome/Toxic epidermal necrolysis
Sangwan Kumari Sushil1, Khanna Neena2, Sharma Namrata3, Agarwal Tushar3, Sharma Arundhati1
1. Laboratory of Cyto-Molecular Genetics, Department of Anatomy, AIIMS, New Delhi 2. Department of Dermatology and Venereology, AIIMS, New Delhi 3. Cornea, Cataract & Refractive Surgery Services, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, AIIMS, New Delhi Presenting author: Sushil Kumari Sangwan Email ID. [email protected] Corresponding author: Arundhati Sharma Email ID. [email protected] Introduction: Stevens-Johnson syndrome (SJS) and Toxic epidermal necrolysis (TEN) are rare but severe dermatological emergencies with high rates of morbidity and mortality. These two clinical entities are described together as epidermal necrolysis (EN). They differ only in their extent of epidermal sloughing viz. SJS is <10%, SJS/TEN overlap is 10-30% and TEN is >30%. Its characteristics are widespread epidermal necrosis of oral, ocular, genital and other mucosal sites. It is classified into 1) acute phase in which patients manifest vesiculobullous lesions of skin and mucosa, severe conjunctivitis and active inflammation and 2) chronic phase in which skin lesions heals up but severe ocular complications persists due to continuous inflammation and corneal epithelial stem cell deficiency. Its etiology is immune-mediated, and is most commonly triggered by the intake of medicines. Survivors of the disorder face trauma not only during the acute phase but also after remission as long-term chronic sequelae. The treatment strategy requires early diagnosis, prompt withdrawal of culprit drug/s, and supportive care with specific therapy in order to reduce severe life-long complications. Aim and Objectives: The aim of the study was to investigate the various aspects underlying the etiology of SJS/TEN. The objectives were to study the clinical profile of patients, the associated findings of genetic and biochemical parameters; and therapeutic outcome in terms of effectiveness. Materials and Methods: 24 subjects diagnosed with acute SJS/TEN were recruited from
Department of Dermatology and Venereology, AIIMS, New Delhi and detailed clinical history and treatment course were noted after taking informed written consent and five ml of peripheral blood samples were drawn. The collected blood samples were subjected to DNA isolation which was utilized to study interleukins and HLA gene polymorphisms and serum separation to check the levels of granulysin and soluble Fas ligand (sFas L). Results: 24 recruited cases (18 females and 6 males) of SJS/TEN, were classified into SJS (25%; 6/24), SJS-TEN overlap (25%; 6/24) and TEN (50%; 12/24) on extent of epidermal detachment. Duration of hospital stay ranged from 8 to 55 days. The triggering drugs noted were non steroidal anti-inflammatory (87.50%; 21/24) followed by antibiotics (66.67%; 16/24), anti epileptics (37.50%; 9/24) and others (37.50%; 9/24). Eruptive lesions of skin were seen in 77.2% (17/24) patients within the first seven days of drug intake, 83.3% (20/24) showed eye related complications during acute phase of the disease along with presence of different co-morbidities in 45% (11/24). Treatment regimen comprised of corticosteroids in 83.3% (20/24) and antimicrobial therapy in 6.6% (4/24). Significantly lower frequency of IL-4 polymorphisms, high frequency of HLA-A*3301 (20.8%), HLA-A*02 (25%), HLA-A*2402 (14.6%) alleles and elevated levels of apoptotic markers - granulysin and sFas L was seen in patients in comparison to controls. Conclusion: The study highlights the usage of corticosteroids and antimicrobial therapy for the effective treatment of patients. It also shows the possible association of IL-4 and HLA-A alleles in the causation of the disease. More number of studies on a larger sample are required which will help in validating the findings of the study and further to assist in improving the patient care
at the earliest phase.
Phd-113
Association of the Genetic variations in monoamine pathway genes with
Alcohol Dependence
Singh Renu, Gupta Ranjan, Grover Tripti, Ambekar Atul, Jain Raka, Sharma Arundhati Laboratory of Cyto-molecular genetics, Department of Anatomy, AIIMS, New Delhi Department of National Drug Dependence and Treatment Center, AIIMS, Ghaziabad Presenting Author: Renu Singh email : [email protected] Corresponding Author: Dr. Arundhati Sharma email: [email protected] Introduction Alcohol, is the most commonly used addictive substance which causes cognitive, behavioral, and physiological changes that lead to alcohol dependence (AD). Its harmful use leads to 3.3 billion deaths/year world-wide and 15 deaths/day in India (WHO 2016). In India, its prevalence is estimated to be ~11% compared to ~16% of the global population (National records bureau and crime 2013).This chronic relapsing brain disorder involves complex interaction between genes and environment as shown by family, twin and adoption studies with about 50–60%heritability. Presence of variations like Single Nucleotide Polymorphisms (SNPs) in the dopamine and serotonin pathway genes has been reported to play important role in alcohol dependence and drug use. These variations affect the functioning of the genes which code for receptors and neurotransmitter that make some individuals more susceptible to alcohol dependence. Aim: To screen for the presence of SNPs of dopamine and serotonin pathway gene and evaluate their role in AD Materials and methods: A total of 100 alcohol dependent males recruited from the Department of Psychiatry (based on DSM-IVR criteria) and 100 healthy males from the general population (controls) formed the study group. Subjects were interviewed using a semi-structured questionnaire and WHO ASSIST, and 5 ml peripheral blood was drawn after taking informed consent. Six SNPs of dopamine pathway DRD4 120bp duplication, DRD3 Ser9Gly, DRD2 Taq1α, COMT V158M, COMT-287A>G, DRD4 -521C/T and four of serotonin pathway HT1B G861C, HTR3B Tyr129Ser, HTR2A T102C and HTR2A A1438G were analyzed by PCR/RFLP. Genotype and allele frequencies were assessed and comparison with alcohol use parameters such as age at first use, duration of alcohol use and liver function tests-GGT, SGOT and SGPT) was done using SPSS v20.0.
Results: Mean age of the patients was (35.7±0.76) years, age at first use (21.8±0.58) years and duration of use was (13.8±0.73) years. Mean SGPT, SGOT and GGT levels were(71.5±5.10;37.2±1.65, 64.3±5.88;43.5±2.29, 3477.518±174.31) in patients and controls. Of the SNPs studied– COMT Val158Met and HTR3B Tyr129Ser showed significant association with AD (p=0.03) while the other SNPs were not significant. Comparison with alcohol use parameters showed an association of HTR1B G861C with liver function (GGT p=0.03) and COMT -287A/G with (SGPT p=0.04). Individuals with HT1B 861C had an increased value of the serum GGT (274.57±98.378) in comparison to those with wild type 861G (75.04±22.064). Similarly, in case of COMT-287A>G, GG genotype carriers showed high SGPT levels (103.75±31.78) as compared to AA carriers (76.23±11.08). HT1B codes for the serotonin receptor and it increases the neurotransmission in reward pathway. Presence of 861C affects the functioning of these receptors and might influence alcohol drinking behaviour. Similarly, COMT gene codes for the enzyme which is involved in dopamine metabolism. Individuals with AA genotype degrade dopamine at normal rates, while GG genotype individuals show slows degradation due to which dopamine remains in the system for more duration which might be responsible for the excess alcohol intake. Conclusion: The present study is indicative of the role of COMT and serotonin gene polymorphisms with AD and suggests that presence of SNPs might influence alcohol related
behavior in dependent individuals.