nephrotoxic effects of uranium in drinking water as detected by urinary protein electrophoresis

1
endogenous and exogenous sources of arsenic and their contribution to hair arsenic levels w:i]l become obv:ous h6 HAIR AS AN INDEX.OF CHROMIUM STATUS AND IMPAIRED GLUCOSE METABOLISM G. Dowe and R. McCurdy, Division of Clinlcal Chemistry, Department of Pathology, Dalhousle University and Victoria General HosFital, Halifax, Nova Scotia, B3H ]vB The potential merits and ]imitations Of ha.ir analyses as an index of chromlum Status will be reviewed in light of the relative inadequacy of more traditlonal biopsy materials. The need far such an index IS further Substantiated by evi- dence indicating that sUboDtimal chromium sutriture may exist in the North Amerclan population with a concomitant incidence of glucose into!lerance associated with decreased levels of glucose tolerance factor (GTF). Difficultles associated with meaningful interpretation ofldata w.i11 be contrasted wi:th the correlatlon of haif chromium to an indlvidua1's known chromium status. The Feasibility of this technieue as a screening test for diabetes melIltus wi11 be examined. 47 NEPHROTOXIC EFFECTS OF URANIUM N DR NK NG WATERAS DETECTED BY URINARY PROTEIN ELECTRoPHbRESIS D.C. Campbell, C. Oymond, M.A. Moss, R.F. McCurdy Dept. of C n cal Chem stry, Dalhous e University, Hal' faxl N.S , BdH 4H7 The purpose of the s~udy was to detect early nephro- toxic e~fects of uranium i.n the drinking water of adult rats. Male Sprague-Dawley rats (8-10 weeks) were Fed uranium, 20 mg per kg body we;ight per day, via gas.tric intubation daily. Twenty-four hour urine specimens were collected over a ~wo week per od and each was concentrated fifty times using Minlcon B-15 concentrators. Electro- phoretic separation of the concen,trate was :carried out using an agarose gel system. The effects, produced by uranium, were manifest by proteinuriawithin as short a ~ime period as one day. Proteinuria was: not only seen as 'ncreased amounts of Albumin but also, and more significantly, as an increase in globular protein, especia]]y in the Beta region of the eIectFophoretic pattern.. ~8 INTERLABORATORY STUDIES IN CLINICAL CHEMISTRY IN CANADA - SUI~Y HIGHLIGHTS OVER A TEN-YEARPERIOD M. W. Weatherburn, Bureau Of Medical Biochemistry, Laboratory Centre for Disease Control, Health and Welfare Canada, Ottawa, KIA OL2 Target values for accuracy have-been assigned by means of CSCC - approved reference methods to haman ~erum samples in nine inter~aboratory studies. Constituents, usually five per study, have included calcium, magnesiam, creatinlne, glucose, sodium, potassium, urate, total protein and cholesterol. Interlaboratory accuracy and precision have been calculated on eachspecimen; intralaboratory accuracy on all specimens within each study. Methods, or method/ instrument systems, have been ranked relative to each other for these three categories of accuracy and precision. Some instraments ranked very well for ear.rain constituen£& yet poorly for others. A number of methods gave accurate results with serum but not with lYophilized materials. Some general Observations common to all constituents are: (i) benefits can be derived from £1ght limits of daily quality control, (2) improved interlaboratory precision can follow from participation in regional quality control programs, (3) potential problems exist in the interpretation of analytical results due ~o (a) wide range of laboratory reference ranges yet no difference in analytical results or (b) differences in analytical results when laboratory reference ranges were similar. h9 DISCRIMINATION BETWEEN OBESE HYPERTENSIVE AND NON- OBESE HYPERTENSIVE PATIENTS ON THE BAS~S OF SERUM LEVE5 OF "PSHUDOCHOLINESTERASE, (PChE). V, M. Prabhakara~, V. ~helan and K. H. Kutty, The Laboratoriesof the General HOspital and Janeway Child Heal:h Centre and Memorial University of Newfoundland, St. John's, Newfoundland, Canada. A large percentage of obese and hypertensive patients have increased serum PChE activity. The objective of this study was to ascertain if any discrimination can be made. between hypar~ens~yeS Wl~h and Wi~hgu~ ObeSity On the basis of Serum PINE, and l~pidS. Prel~minary c~se studz o~ I0 patients with h~ertensi0n iS presented, In non-0hese h~erltensiVe~'~5~,serum ~I~ChE; showed @ante Of 2283~36~6 ~O/:L {mean 5005 Z 11 SEMi, , " ~ri~iyeerideS a range Of 75:-= l:Sl mE%; (~iS +- 2 ~44) and che~eStez01 ca rang e of t72 - 2~3 mg~ (205 -+ 2.45), By' contrast, ~n dbes:e h~ertefis£ve~s.~se~ PChHh~s a range 3550~6777 IU/L (mean 5232 - 15.2 S£24), • rigl),eer&des ~ range 0f:i[35 - 120S rag% (476 • 9,2)and chd~esterol a. range of 252 - 500 mg~ (273 +~3.19. Normal range on eu~ methods: f o r pChE i s 2400 ~ 3400 IU/L, tr~glycerides 74 - 177 mg~ and Cholesterol 150 - 250 mg~. Ne ~he~efore pro~se that serum P~E:has a good diserim2natiue, value in ldentify~ng n0n~obese, ner~- l~pemic ~ypertensives from obese,, h~erl~pemic h~ertensives. 510: URINARY: PRO~FEINS iN' BREAST C~NCER C.L. Cousins and S.D~ Rice, Department :of Pathology; Div;~ of c ii:nic@l Chemi.stry, Vic~oF!a ~enePa:i'Hosp]tai, Halifax, Nova Scot.is, B3H iV8 C0ncentra~ed humar urinary p..ro~eins fr0mPati.ents w!~h breast cancer and a control grouP were sep@,ratedby using high reso]utlon two-d!mensionaI e ectr0phoresi:s: The ~esuIts of this preIiminarY :study show basic similarities in a:]] patients sinre thece a:~e flve charaCterlstic land- markS in the e]ectropho~etlC patterps. However,in six out of the e]even cancer patients here is a l'i:ne of Unresol.ved protein/s that travels jus~ below and to the right 6f ~1 - acld glycoprotein which is not present in the. seven patients of the control group. This dlf~erence may be asS0ciat~d with the ~umor and its appearance may be :dependent on the type, stage and grade of the tumor. This protelh or pr0teins may have clini:cal significance as a pbtenti:al mark" er protein for breast cancer. 51 EARLY TUBULAR LESIONS I:N :RAT KDNEYS AFTER URANIUMEXPOSURE K.E, Keller, A. Trillo, M.A, Mess, R.F. MeOutdy~ D.C. campbell:, Department of Pathology, DBIhousI:euniversity Halifax, NOVb ;SdOtia and V:ictoria General Hospltal, :Halifax Nova Sc0tcia, B3H IV8 Thi:s is a report Of changes seen by l],ghtand e!ecfron microscopy in rat kidneys after uranium i:nj:ury. During preliminary Studies. a group of male Spragu e Dawley rats were dosed W;i'th u~anYI nltra:te so]utlon vla gastrlC tube, over a tWO week period. The morpho :og cal changes during th e injury and recovery phase have been co'rrela~ed with p01yur-la and pKoteinuria occurring in these anima.l:s Our Sl;Udy is part of an investiga:tion ~o estabiish the tOXiC effects o:f uranium in d:rl.nking water., correlating morPhologlca'! and::chemlca! indicators pf injury and pre- y:in:log toxicological data relevant tO human drinking water gu:ideiines for uranium., EXPERIMENTAL PATHOLOGY POSTERS 52: THE:ROLE OF PMNs. IN THE PATHOGENESIS OF SlLICOTiG FIBROSIS. D.H, .Bowden and I,Y.R. Ad~son, .Dept. Of Pathology, U~vers~gy Of Man±toba, Winnipeg, ManitOba R3E OW3 Silicosls iS usually: attributed to flb~oblast st]mOlar]on by secret]otis Of damaged alveolar maerophag~s (AMg, The role of FMNS and elf continuing dell ~njo~ in the pathogenes~s now studled. MiCe, injec:ted in~ra~racheally ~th 2~ SiO2~ reCelv~ ed 3H thymldifie I hour before death at intervals tO 20 weeks. cellular populatiOnS and lySosomal content of lavage fluidS: were correlated ~th: morphology, DNA Synthesis and COllagen C0ntent of tNe Iong. The Iniglal response invo1~ed rapid PMN and AM recruitment ~0 ~he alveoll. Foea~ Type. 1 cell damage was rapld~y repaired by Type 2 tell pr011fe~a~i0n. ~l:though PMN and numbers dropped' after a ~ew days, ~h~y n~er reached Control [evils and rbse again after 8 weeks. Disrupt~ 1onof bdeh PMNs and AMS w~th Silica WaS observed in legated Cell pellets<. G[uCosamin~dase and glucur0[nidaS~ levels were~ a~ways h~gher than ~ontr01s thr0ugh0u= ~he s~udy. Labe1~ng Studies :shOwed contlnui~g ~le~ted :D~ synthes&S by: in~er~ Stit~al cells tO 20 weeks; Increased c0[~agen synthesls was found from 4 weeks onward, some free Pa~C~e~aS ~rossgd TM~g i epithelia! cells: and a few: silica ~art~cles were fqund interstitial macr.ophages, The res~s ~ ind±ca~e ~hat ~e0ntlnu~n8 injury £0 and ~s in al~eeil and ~n the ~th CLINICAL BIOCHEMISTRY, VOLUME 17, FEBRUARY 1984 207

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endogenous and exogenous sources o f arsenic and t h e i r c o n t r i b u t i o n to ha i r arsenic leve ls w:i ] l become obv:ous

h6 HAIR AS AN INDEX.OF CHROMIUM STATUS AND IMPAIRED GLUCOSE METABOLISM

G. Dowe and R. McCurdy, D iv i s ion o f C l i n l ca l Chemist ry , Department of Pathology, Dalhousle University and V i c t o r i a General HosFital, Halifax, Nova Scotia, B3H ]vB

The potential merits and ] imitations Of ha.ir analyses as an index of chromlum Status wi l l be reviewed in l ight o f the relat ive inadequacy of more tradit lonal biopsy materials. The need far such an index IS further Substantiated by evi- dence indicating that sUboDtimal chromium sutr i ture may exist in the North Amerclan population with a concomitant incidence of glucose into!lerance associated with decreased levels of glucose tolerance factor (GTF). D i f f icu l t les associated with meaningful interpretation ofldata w.i11 be contrasted wi:th the correlatlon of haif chromium to an indlvidua1's known chromium status. The Feasibi l i ty of this technieue as a screening test for diabetes melIltus wi11 be examined.

47 NEPHROTOXIC EFFECTS OF URANIUM N DR NK NG WATER AS DETECTED BY

URINARY PROTEIN ELECTRoPHbRESIS

D.C. Campbell, C. Oymond, M.A. Moss, R.F. McCurdy Dept. of C n cal Chem stry, Dalhous e University, Hal' faxl N.S , BdH 4H7

The purpose of the s~udy was to detect early nephro- toxic e~fects of uranium i.n the drinking water of adult rats. Male Sprague-Dawley rats (8-10 weeks) were Fed uranium, 20 mg per kg body we;ight per day, via gas.tric intubation daily. Twenty-four hour urine specimens were collected over a ~wo week per od and each was concentrated f i f t y times using Minlcon B-15 concentrators. Electro- phoretic separation of the concen,trate was :carried out using an agarose gel system. The ef fects, produced by uranium, were manifest by proteinur iawith in as short a ~ime period as one day. Proteinuria was: not only seen as 'ncreased amounts of Albumin but also, and more signi f icant ly, as an increase in globular protein, especia]]y in the Beta region of the eIectFophoretic pattern..

~8 INTERLABORATORY STUDIES IN CLINICAL CHEMISTRY IN CANADA - SUI~Y HIGHLIGHTS OVER A TEN-YEARPERIOD

M. W. Weatherburn, Bureau Of Medical Biochemistry, Laboratory Centre for Disease Control, Health and Welfare Canada, Ottawa, KIA OL2

Target values for accuracy have-been assigned by means of CSCC - approved reference methods to haman ~erum samples in nine inter~aboratory studies. Constituents, usually five per study, have included calcium, magnesiam, creatinlne, glucose, sodium, potassium, urate, total protein and cholesterol. Interlaboratory accuracy and precision have been calculated on eachspecimen; intralaboratory accuracy on all specimens within each study. Methods, or method/ instrument systems, have been ranked relative to each other for these three categories of accuracy and precision.

Some instraments ranked very well for ear.rain constituen£& yet poorly for others. A number of methods gave accurate results with serum but not with lYophilized materials. Some general Observations common to all constituents are: (i) benefits can be derived from £1ght limits of daily quality control, (2) improved inter laboratory precision can follow from participation in regional quality control programs, (3) potential problems exist in the interpretation of analytical results due ~o (a) wide range of laboratory reference ranges yet no difference in analytical results or (b) differences in analytical results when laboratory reference ranges were similar.

h9 DISCRIMINATION BETWEEN OBESE HYPERTENSIVE AND NON- OBESE HYPERTENSIVE PATIENTS ON THE BAS~S OF SERUM LEVE5 OF "PSHUDOCHOLINESTERASE, (PChE).

V, M. Prabhakara~, V. ~helan and K. H. Kutty, The Laboratoriesof the General HOspital and Janeway Child Heal:h Centre and Memorial University of Newfoundland, St. John's, Newfoundland, Canada.

A large percentage of obese and hypertensive patients have increased serum PChE activity. The objective of this study was to ascertain if any discrimination can be

made. between hypar~ens~yeS Wl~h and Wi~hgu~ ObeSity On the basis of Serum PINE, and l~pidS. Prel~minary c~se studz o~ I0 patients with h~ertensi0n iS presented,

In non-0hese h~erl tensiVe~'~5~,serum ~I~ChE; showed @ante Of 2283~36~6 ~O/:L {mean 5005 Z 11 SEMi, , " ~ri~iyeerideS a range Of 75:-= l:Sl mE%; (~iS +- 2 ~44) and che~eStez01 ca rang e o f t72 - 2~3 mg~ (205 -+ 2.45) , By' contrast, ~n dbes:e h~ertef is£ve~s.~se~ PChHh~s a range e£ 3550~6777 IU/L (mean 5232 - 15 .2 S£24), • r igl) ,eer&des ~ r a n g e 0f:i[35 - 120S rag% (476 • 9 , 2 ) a n d chd~esterol a. range of 252 - 500 mg~ (273 +~3.19. Normal range on eu~ methods: f o r pChE i s 2400 ~ 3400 IU/L, tr~glycerides 74 - 177 mg~ and Cholesterol 150 - 250 mg~.

Ne ~he~efore p r o ~ s e t h a t serum P~E:has a good diserim2natiue, value in ldentify~ng n0n~obese, ner~- l~pemic ~ypertensives from obese,, h~erl~pemic h~ertensives.

510: URINARY: PRO~FEINS iN' BREAST C~NCER C.L. Cousins and S.D~ Rice, Department :of Pathology; Div;~ o f c ii:nic@l Chemi.stry, Vic~oF!a ~enePa:i'Hosp]tai, Halifax, Nova Scot.is, B3H iV8

C0ncentra~ed humar urinary p..ro~eins fr0mPati.ents w!~h breast cancer and a control grouP were sep@,rated by using high reso]utlon two-d!mensionaI e ect r0phores i :s : The ~esuIts of th is preIiminar Y :study show basic s imi lar i t ies in a:]] patients sinre thece a:~e f lve charaCterlstic land- markS in the e]ectropho~etlC patterps. However, in six out of the e]even cancer patients here is a l'i:ne of Unresol.ved protein/s that travels jus~ below and to the r ight 6f ~1 - acld glycoprotein which is not present in the. seven patients o f the control group. This dlf~erence may be asS0ciat~d with the ~umor and its appearance may be :dependent on the type, stage and grade of the tumor. This protelh or pr0 te ins may have c l in i :ca l s i g n i f i c a n c e as a pb ten t i :a l mark" er protein for breast cancer.

51 EARLY TUBULAR LESIONS I:N :RAT KDNEYS AFTER URANIUM EXPOSURE

K.E, Keller, A. T r i l l o , M.A, Mess, R.F. MeOutdy~ D.C. campbell:, Department of Pathology, DBIhousI:e university Halifax, NOVb ;SdOtia and V:ictoria General Hospltal, :Halifax Nova Sc0tcia, B3H IV8

Thi:s is a report Of changes seen by l],ght and e!ecfron microscopy in ra t k idneys a f t e r uranium i :nj :ury. During preliminary Studies. a group of male Spragu e Dawley rats were dosed W;i'th u~anYI nltra:te so]utlon vla gastrlC tube, over a tWO week period.

The morpho :og cal changes during th e injury and recovery phase have been co'rrela~ed with p01yur-la and pKoteinuria occurring in these anima.l:s

Our Sl;Udy is part o f an investiga:tion ~o estabiish the tOXiC ef fec ts o:f uranium in d:rl.nking water., correlating morPhologlca'! and::chemlca! indicators pf injury and pre- y:in:log toxicological data relevant tO human drinking water gu:ideiines for uranium.,

EXPERIMENTAL PATHOLOGY POSTERS

52: THE:ROLE OF PMNs. IN THE PATHOGENESIS OF SlLICOTiG FIBROSIS.

D.H, .Bowden and I,Y.R. Ad~son, .Dept. Of Pathology, U~vers~gy Of Man±toba, Winnipeg, ManitOba R3E OW3

Silicosls iS usually: attributed to flb~oblas t st]mOlar]on by secret]otis Of damaged alveolar maerophag~s (AMg, The role of FMNS and elf continuing dell ~njo~ in the pathogenes~s now studled. MiCe, injec:ted in~ra~racheally ~th 2~ SiO2~ reCelv~ ed 3H thymldifie I hour before death at intervals tO 20 weeks. cellular populatiOnS and lySosomal content of lavage fluidS: were correlated ~th: morphology, DNA Synthesis and COllagen C0ntent of tNe Iong. The Iniglal response invo1~ed rapid PMN and AM recruitment ~0 ~he alveoll. Foea~ Type. 1 cell damage was rapld~y repaired by Type 2 tell pr011fe~a~i0n. ~l:though PMN and numbers dropped' after a ~ew days, ~h~y n~er reached Control [evils and rbse again after 8 weeks. Disrupt ~ 1onof bdeh PMNs and AMS w~th Silica WaS observe d in legated Cell pellets<. G[uCosamin~dase and glucur0[nidaS~ levels were ~ a~ways h~gher than ~ontr01s thr0ugh0u= ~he s~udy. Labe1~ng Studies :shOwed contlnui~g ~le~ted :D~ synthes&S by: in~er~ Stit~al cells tO 20 weeks; Increased c0[~age n synthesl s was found from 4 weeks onward, some free Pa~C~e~aS ~rossgd TM~g i epithelia! cells: an d a few: silica ~art~cles were fqund interstitial macr.ophages, The res~s ~ ind±ca~e ~hat ~e0ntlnu~n8 injury £0 and ~s in al~eeil and ~n the ~th

CLINICAL BIOCHEMISTRY, VOLUME 17, FEBRUARY 1984 207