Estrogen receptor-alpha deficiency attenuates autoimmune disease in (NZB x NZW)F1 mice.

Estrogens promote lupus in humans and some mouse models of this disease. Nonetheless, little is known about the role of estrogen receptors in lupus pathogenesis. Here, we report that in females on the lupus-prone (NZB x NZW)F(1) background, disruption of estrogen receptor-alpha (ER alpha or Esr1) attenuated glomerulonephritis and increased survival. ER alpha deficiency also retarded development of anti-histone/DNA antibodies, suggesting that ER alpha promotes loss of immunologic tolerance. Furthermore, ER alpha deficiency in (NZB x NZW)F(1) females attenuated the subsequent development of anti-double-stranded DNA (dsDNA) IgG antibodies, which are associated with glomerulonephritis in this model. We provide evidence that ER alpha may promote lupus, at least in part, by inducing interferon-gamma, an estrogen-regulated cytokine that impacts this disease. ER alpha deficiency in (NZB x NZW)F(1) males increased survival and reduced anti-dsDNA antibodies, suggesting that ER alpha also modulates lupus in males. These studies demonstrate that ER alpha, rather than ER beta, plays a major role in regulating autoimmunity in (NZB x NZW)F(1) mice. Furthermore, our results suggest for the first time that ER alpha promotes lupus, at least in part, by impacting the initial loss of tolerance. These data suggest that targeted therapy disrupting ER alpha, most likely within the immune system, may be effective in the prevention and/or treatment of lupus.

Glomerular structure in IDDM women with low glomerular filtration rate and normal urinary albumin excretion.

Eight women with insulin-dependent diabetes mellitus (IDDM) with low creatinine clearance rate (CCR) and normal urinary albumin excretion (UAE) were compared with three other groups of diabetic women: 19 with normal creatinine clearance rate (CCR) and UAE, 7 with normal CCR and microalbuminuria, and 7 with low CCR and microalbuminuria. The four groups were similar in age, duration of diabetes, HbA1, incidence of urinary tract infection, prevalence of bladder neuropathy, and urinary urea nitrogen excretion rate. The prevalence of hypertension was similar among the groups, although mean arterial pressure was higher in the low CCR and microalbuminuria group. Renal area index was lower in the low CCR and normal UAE groups than in the other groups of diabetic patients, but was not different from normal. Morphometric measures of mesangial expansion and estimates of arteriolar hyalinosis and global glomerulosclerosis were increased to a similar degree in the low CCR and normal UAE, normal CCR and microalbuminuria, and low CCR and microalbuminuria groups compared with the group without abnormalities of renal function. Therefore, it is likely that diabetic glomerulopathy is, at least in part, responsible for the loss of glomerular filtration rate seen in the low CCR and normal UAE patients. Thus, the definition of incipient nephropathy may have to be expanded beyond the concept of microalbuminuria if longitudinal study of such patients reveals an increased risk of the subsequent development of overt nephropathy. Finally, screening for diabetic kidney disease among IDDM patients should include determination of glomerular filtration rate and measurement of UAE and blood pressure, especially among women.

5-thio-D-glucose elevates renal transforming growth factor beta-1 at a dose that does not prevent streptozocine diabetes in rats.

Studies of early nephropathy in streptozocin (STZ)-treated rats are complicated by the nephrotoxicity of this agent. Inhibitors of the diabetogenic actions of STZ have been described, but their effects on the kidney have not been assessed. This study examined the effects of one agent, 5-thio-D-glucose (5TG) on renal hypertrophy and transforming growth factor beta1 (TGF-beta1). Forty male Sprague Dawley rats were divided into four groups: saline controls (SC), 5TG alone, 5TG + STZ, and STZ. After 2 weeks of observation, urine, plasma, and kidneys were studied. Nine of 10 STZ rats were diabetic at the time of euthanasia, as were 5 of 10 5TG + STZ animals. Both tissue levels of messenger RNA and protein for active and total TGF-beta1 were elevated in STZ and 5TG-STZ animals compared with SC. 5TG also elevated mRNA and produced protein levels intermediate to the other groups. 5TG plus STZ is an unacceptable control for nephropathy studies in STZ diabetes, both because of lack of efficacy at the dose studied and the induction of TGF-beta1 by 5TG. 5TG may yet prove of value in studying control of renal TGF-beta1 expression and excretion.

Long-term furosemide treatment in the normal rat: dissociation of glomerular hypertrophy and glomerulosclerosis.

Furosemide treatment produces glomerular hypertrophy and augments glomerular capillary hydraulic pressure in the normal rat. Similar processes have been implicated in the progression of glomerulosclerosis (GS). Whereas prior experiments with furosemide treatment of 6 to 8 weeks duration have produced no detrimental effects on renal function or structure, the effects of more prolonged treatment are unknown. Male Munich-Wistar rats were pair fed with or without furosemide, 40 mg/d, from the time of weaning through 10 months of age. At selected time points, 24-hour urine collections were obtained for total protein and volume determination. At the end of the study, light and electron microscopic morphometric studies were performed. Renal cortical hypertrophy and glomerular hypertrophy were sustained throughout the 9 months of treatment in the group receiving furosemide. The cortical interstitial area was increased in the furosemide group, but this did not appear to be the result of fibrosis. Proximal and distal tubule diameter were unaffected by treatment. No differences in GS or glomerular ultrastructure were shown. This study provides no evidence of detrimental glomerular effects of furosemide in normal animals. Further studies of furosemide treatment under conditions of preexisting renal pathological conditions are warranted to confirm the safety of this treatment in situations analogous to those seen in the clinical setting. Interstitial expansion also warrants further study in this setting.

Outcome of dialysis for acute renal failure in pediatric bone marrow transplant patients.

We have reviewed the clinical course of 30 pediatric bone marrow transplant (BMT) recipients requiring dialysis for acute renal failure early after BMT. Patients requiring dialysis were not significantly different from the general pediatric BMT population except for: (1) a greater proportion of neuroblastoma patients in the dialysis group, and (2) fewer autologous and more unrelated BMT donors in the dialysis group. Twenty-three patients (77%) died without recovering renal function 1-72 days (mean 12 days) after dialysis was begun. Sepsis was the most commonly cited cause of renal failure and death in these patients. Seven patients (23%) recovered sufficient renal function to stop dialysis; all long-term survivors were in this group. Factors at the onset of dialysis associated with persistent renal failure were weight gain of > or = 10% of baseline body weight, requirement of three or more drugs for blood pressure support and hyperbilirubinemia. Although acute renal failure requiring dialysis is an ominous development following BMT, recovery of renal function is possible with aggressive supportive care.

Furosemide treatment, angiotensin II, and renal growth and development in the rat.

Diuretics are commonly used to treat a variety of conditions, including progressive nephropathies, cardiovascular, and pulmonary diseases. Such treatment stimulates the production of angiotensin II, an important mediator of renal growth which may accelerate progressive glomerulosclerosis. To further study the effects of diuretic treatment on normal renal growth and development, weanling male Munich-Wistar rats received no drug, enalapril, furosemide, or both drugs for 6 wk; morphometric studies were then performed using standard light and electron microscopic techniques. Plasma renin activity was elevated by furosemide treatment. Cortical tubular growth was stimulated in rats receiving furosemide or both drugs; enalapril did not affect cortical tubular growth when compared with untreated animals. Glomerular volume was increased in furosemide-treated animals, primarily due to an increase in the proportion of mesangial cells, whereas enalapril decreased glomerular volume. Furosemide also increased the filtration surface area per glomerulus whereas enalapril decreased it. Concurrent enalapril treatment blocked the furosemide-induced changes in filtration surface area as well as attenuating overall glomerular and mesangial growth. Glomerular changes correlated with plasma renin activity. Furosemide stimulated glomerular growth, especially of mesangial cells, probably via stimulation of AngII production. Given the relationship of mesangial cell growth and progressive renal disease, diuretic therapy could thus accelerate glomerulosclerosis. Cortical tubular growth also increased with furosemide; however, enalapril had no effect, so factors besides angiotensin II appeared to modulate tubulointerstitial growth in these animals.

Determination of mean glomerular volume in nephrectomy specimens.

BACKGROUND: Mean glomerular volume (VG) determined by Cavalieri, maximal profile area (MPA), and Weibel-Gomez methods correlated in paraffin-embedded needle biopsies; the Disector method did not correlate, probably because of sampling limitations. The relationship of these methods in other types of specimens has not been established. EXPERIMENTAL DESIGN: Sections of whole kidneys from 19 rats, 10 treated with furosemide to increase VG, were immersion-fixed, embedded in glycol methacrylate, serially sectioned at 5 microns through 150 microns of tissue, and stained with Jones' silver stain. VG was determined by the Cavalieri, MPA, Disector, and Weibel-Gomez methods with three samples from each animal to assess the correlation, agreement, and repeatability of each of these methods. Methods were considered to correlate if p < 0.01 on linear regression analysis and to agree if the line of identity (x = y) fell within the 95% confidence interval for the slope of the regression curve. Repeatability was assessed with the coefficient of variation for each technique. RESULTS: Other methods correlated with the Cavalieri method (MPA r = 0.86, Disector r = 0.66, Weibel-Gomez r = 0.87), although none of these methods agreed with Cavalieri VG. These results were similar when treated animals, untreated animals, and all animals were analyzed. Cavalieri and MPA methods showed 7% coefficient of variation with repeated samples; the Disector method showed higher variation (12%), and Weibel-Gomez showed significantly higher variation when compared with the other methods (25%). CONCLUSIONS: MPA, Weibel-Gomez, and Disector methods all provide estimates of VG that correlate with Cavalieri VG; however, none gives the same value. Reasons for these discrepancies may include assumptions regarding glomerular shape and size distribution inherent in the various methods and the experimental conditions. Tissue availability will often dictate VG method, and consistency of method within an experiment is critical.

Age of onset of streptozocin diabetes determines the renal structural response in the rat.

Prepubertal years of insulin-dependent diabetes mellitus are protected from the nephropathic effects of this disease, yet this effect of immaturity has not been investigated in an animal model. Male Munich-Wistar rats were made diabetic with streptozocin at two ages: weanling (approximately 4 wk) and pubescent (approximately 10 wk). After 5 wk of untreated diabetes, weanling diabetic animals showed relatively greater growth of the medulla, whereas relative proportions of these areas were constant in the older animals. Glomerular volume increased by approximately 35% in older diabetic animals, but no glomerular enlargement was demonstrated in weanling rats with diabetes. Glomerular ultrastructure was not significantly altered during the short course of this study. The renal structural response to diabetes is age-dependent in the rat, with prepubertal animals protected from glomerular hypertrophy. Longer studies are needed to see if these differences will eventually parallel those demonstrated in patients with onset of diabetes before and after puberty. This model may ultimately prove to be valuable in determining the mechanism via which prepubertal kidneys are protected from the nephropathic effects of insulin-dependent diabetes mellitus.

Estimation of glomerular volume: a comparison of four methods.

Methods for estimating glomerular volume were compared in Zenker-fixed, paraffin-embedded biopsies from 10 patients with insulin-dependent diabetes mellitus and 6 normal kidney donors. Two methods of measurement of individual glomerular volumes were used: the Cavalieri method (considered the "gold standard") and the maximal profile area (MPA) method. Also studied were the method of Weibel and Gomez and a method based on the disector principle; both estimate mean volume (VG). MPA and Cavalieri showed strong correlation (r = 0.93; P less than 0.001), although the MPA method consistently overestimated the true volume; six glomeruli were necessary for a reliable estimate of VG. The disector method did not correlate with VG determined by Cavalieri. Weibel-Gomez did correlate with Cavalieri (r = 0.68; P less than 0.05), but overestimated VG. At least 15 profiles were needed to provide a dependable estimate of VG by Weibel-Gomez. The Cavalieri, MPA, and Weibel-Gomez methods all can provide reliable estimates of VG, the latter two with appropriate correction factors. The individual glomerular volume methods, while more time consuming, provide information on variation and distribution of the glomerular population and are the methods of choice for studies of glomerular volume.

Chronic administration of furosemide augments renal weight and glomerular capillary pressure in normal rats.

Angiotensin II (ANG II) is believed to promote progressive renal injury via augmented glomerular capillary hydraulic pressure (PGC). Acute volume reduction secondary to diuretic administration increases circulating ANG II and augments PGC, yet the hemodynamic effects of sustained diuretic administration are unknown. Therefore, glomerular micropuncture studies were performed in male Munich-Wistar rats after 6-8 wk of treatment with daily furosemide (F, 40 mg/day), furosemide plus the AT1 receptor antagonist, losartan (F + L, 5 mg/day), or no therapy (C, control). Renal weight was increased in F rats (1.23 +/- 0.7 g) vs. C (1.00 +/- 0.06 g) or F + L (0.97 +/- 0.01 g). In addition, PGC was elevated in F animals (52.1 +/- 1.5 mmHg) vs. C (43.7 +/- 1.5) or F + L-treated rats (41.3 +/- 1.7). F-treated rats were also characterized by a relative increase in efferent arteriolar resistance and filtration fraction. The latter was markedly attenuated in F + L-treated animals. Collectively, these findings are consistent with an ANG II-mediated alteration in intrarenal hemodynamics. In contrast to acute volume manipulations, however, chronic furosemide augmented renal growth, whereas losartan administration completely arrested this phenomenon. Further studies are warranted to determine whether the hemodynamic and growth adaptations elicited by chronic F administration induce or accelerate renal injury.

Streptozocin diabetes elevates all isoforms of TGF-beta in the rat kidney.

Transforming growth factor beta (TGF-beta) is a major promoter of diabetic nephropathy. While TGF-beta1 is the most abundant renal isoform, types 2 and 3 are present as well and have identical in vitro effects. Whole kidney extracts were studied 2 weeks after induction of streptozocin diabetes and in control rats. Mean glomerular area was 25% greater in the diabetic animals. TGF-beta1 showed a 2-fold increase in message with a 3-fold increase in protein. TGF-beta2 mRNA increased approximately 6% while its protein doubled. TGF-beta-message increased by 25%, producing a 35% increase in its protein. TGF-beta-inducible gene H3 mRNA was increased 35% in the diabetic animals, consistent with increased activity of this growth factor. All isoforms of TGF-beta are increased in the diabetic rat kidney. Future studies need to address the specific role that each isoform plays in diabetic nephropathy as well as the impact of therapies on each isoform.

Sex differences in the renal transforming growth factor-beta 1 system after puberty.

Transforming growth factor-beta 1 (TGF-beta 1) has been implicated in many progressive kidney diseases. The present study examines this growth factor during the pubertal and early adult periods. Mixed-sex Munich-Wistar rat kidneys were obtained on selected days of life from birth through 6 months of age. A survey of the TGF-beta 1 system was performed, and then a second experiment focused on cortex and medulla from both sexes at 6 weeks and 16 weeks of age. Reverse transcription polymerase chain reaction for TGF-beta 1 and TGF-beta inducible gene H3 (beta IG-H3) was performed. Active and total levels of protein for TGF-beta 1 were isolated from tissue. Active levels of TGF-beta 1 were somewhat lower in older than in younger animals, without sex differences. beta IG-H3 levels were similar. At 16 weeks females had levels of total growth factor approximately threefold greater than males, while adult males appeared to activate the growth factor much more efficiently. These findings suggest that activation of TGF-beta 1 becomes more efficient following puberty in the male rat, while females appear to have reduced activation efficiency compensated by increased total growth factor. These differences may help explain the deterioration at puberty and sexual dimorphism noted with some progressive nephropathies.

Renal interstitial expansion in insulin-dependent diabetes mellitus.

Eighty-four patients with insulin-dependent diabetes mellitus had studies of renal function and quantitative renal morphometry including mesangial volume fraction (Vvmes/glom), index of arteriolar hyalinosis, percentage of globally sclerosed glomeruli (%GS), and interstitial volume fraction for total renal cortex (Vvint/T). There was significant correlation among these four parameters, and all four structural parameters correlated with glomerular filtration rate and the log of urinary albumin excretion. Stepwise multiple regression analysis showed that Vvmes/glom and Vvint/T were additive, suggesting that they are partially independent. Arteriolar hyalinosis and %GS did not improve the correlations further. We hypothesize that Vvmes/glom, Vvint/T, arteriolar hyalinosis, and %GS represent multiple but probably interrelated pathologic mechanisms leading to the functional disturbances of diabetic nephropathy. Longitudinal studies of patients with diabetes and studies of patients with diseases producing interstitial expansion in the absence of glomerular disease may help clarify the independent role of interstitial expansion in the kidney disease of diabetes mellitus.

Steroid-dependent nephrotic syndrome following renal transplantation for congenital nephrotic syndrome.

A boy developed recurrent steroid-responsive nephrotic syndrome following renal transplantation for congenital nephrotic syndrome. The first episode was associated with mild tubulointerstitial rejection on kidney biopsy. Subsequent episodes showed normal histology by light microscopy and epithelial foot process fusion on electron microscopy, consistent with minimal change nephrotic syndrome. Serum analysis for soluble immune response suppressor was negative pre-nephrectomy, positive during each bout of nephrotic syndrome, and negative during each remission. This case represents de novo occurrence of steroid-sensitive minimal change nephrotic syndrome following renal transplantation for congenital nephrotic syndrome. We stress the need for histological examination of the renal allograft to diagnose rejection, recurrent disease, or de novo disease.

Glomerular structural and functional changes in a high-fat diet mouse model of early-stage Type 2 diabetes.

AIMS/HYPOTHESIS: Type 2 diabetes often results in diabetic nephropathy, which is preceded by an elevated glomerular filtration rate (GFR). This study was designed to develop a mouse model of Type 2 diabetes and to elucidate the glomerular events in the early stages of diabetic nephropathy. METHODS: Four-week-old mice were fed a normal or high-fat (42% of total calories from fat) diet, and body weight, blood glucose, insulin, leptin, lipids and GFR were monitored from 9 to 21 weeks or longer after the feeding programme. Mesangial cell dedifferentiation was accessed by alpha-smooth muscle actin staining. Glomerular hypertrophy was determined using image analysis with haematoxylin-eosin staining. Matrix deposition was determined by type IV collagen staining. RESULTS: After 9 weeks, mice fed a high-fat diet weighed more than mice fed a normal diet (30.5+/-1.2 vs 22.3+/-0.5 g, p<0.05), and mice fed a high-fat diet were hyperinsulinaemic (283.9+/-69.7 vs 102.9+/-36.4 pmol/l, p<0.05), hyperglycaemic (8.0+/-0.6 vs 6.5+/-0.2 mmol/l, p<0.05) and their leptin levels were increased six-fold (1.48+/-0.45 vs 0.25+/-0.03 ng/ml, p<0.05). After 13 weeks, mice fed a high-fat diet showed hyperfiltration (GFR; 440+/-60 vs 210+/-10 microl/min, p<0.05). During the early stages of diabetic nephropathy, mesangial cell dedifferentiation was evident, shown by increased expression of alpha-smooth muscle actin in the glomeruli. After 9 weeks, mice fed a high-fat diet already demonstrated increased type IV collagen deposition. After 13 weeks, they developed enlarged glomerular tufts compared with those of their age-matched controls. CONCLUSIONS/INTERPRETATION: The results of this study suggest that collagen IV deposition precedes the hyperfiltration and enlargement of glomeruli in early-stage diabetic nephropathy. Dedifferentiation of mesangial cells may be associated with collagen IV deposition.

Puberty permits increased expression of renal transforming growth factor-beta1 in experimental diabetes.

Prepubertal years of diabetes mellitus are relatively protected from clinical manifestations of nephropathy. Transforming growth factor-beta1 (TGF-beta1) is a major mediator of diabetic kidney disease. Its renal expression, translation, and activation change with sexual maturation in the normal rat. The role of TGF-beta1 in postpubertal susceptibility to diabetic renal hypertrophy was addressed in the present study. Male Sprague- Dawley rats were given streptozocin at 4 weeks of age (weanling) or 14 weeks of age (mature) and treated with insulin to maintain blood glucose levels between 300 and 500 mg/dl. Nondiabetic controls received saline. After 6 weeks with ad libitum food and water, kidneys were snap-frozen for measurement of TGF-beta1 protein and mRNA. As in previous studies, diabetic renal hypertrophy was blunted in weanling animals compared with mature rats. Message for TGF-beta1 was not significantly increased in weanling animals [102 (9)% [mean (SEM)] in nondiabetic controls versus 117 (10)% in diabetic rats; P=0.91], while it was significantly increased in mature diabetic animals [100 (7)% vs. 146 (11)%; P=0.01]. Immunohistochemistry revealed focal increases in glomerular staining in mature but not weanling diabetic rats. Differences in the control of the renal TGF-beta system may explain the permissive role of puberty in the manifestations of diabetic kidney disease.

Group B coxsackievirus myocarditis and pancreatitis: connection between viral virulence phenotypes in mice.

The group B coxsackieviruses (CVB) induce experimental pancreatitis and myocarditis in mice and are established agents of human myocarditis, especially in children. We tested the hypothesis that the development of CVB-induced myocarditis is linked to CVB-induced pancreatitis by studying the replication of different CVB strains in mice. Eight of nine genotypically different type 3 CVB (CVB3) strains induced acute pancreatitis in mice; of these, three viruses also induced acute myocarditis. One CVB3 strain was avirulent for both organs. Myocarditis was not observed in the absence of pancreatitis. The results obtained by inoculation of mice with strains of other CVB serotypes were consistent with these data. Infectious virus titers were measured in serum, pancreas, and heart as a function of time after inoculation of mice with three CVB3 strains. Each strain was representative of one of the three viral virulence phenotypes: avirulent, pancreovirulent only, and cardiovirulent. All strains replicated well and persisted in the pancreas through 8 days post-inoculation, but the cardiovirulent CVB3 strain tended to replicate to higher titer earlier and persist longer in sera, pancreatic, and cardiac tissues than the noncardiovirulent strains. Replication of the CVB3 strains were studied in two human pancreatic tumor lines and in primary human endothelial cell cultures derived from cardiac artery. Cardiovirulent strains, both individually and as a group, tended to replicate to titers as high as, or higher than, noncardiovirulent strains did in cell culture. The data are consistent with the possibility of an etiologic link between CVB-induced pancreatic and heart disease.

Effect of epidermal growth factor in the rat 5/6 renal ablation model.

Acute administration of epidermal growth factor (EGF) has been shown to promote recovery from ischemic and nephrotoxic acute renal failure in vivo. The question of whether chronic subcutaneous administration of EGF (19.1 micrograms/day for 3 or 6 wk) could alter the course of chronic renal failure in rats subjected to 5/6 nephrectomy was studied. By week 6, there was no difference in renal function, as assessed by animal survival, BUN, urea and inulin clearances, proteinuria, renal morphometry, or renal size, between EGF- and vehicle-treated rats. This suggests that chronic renal insufficiency differs from acute tubular injury in its sensitivity to exogenous EGF. Unexpectedly, EGF significantly attenuated the rise in systolic blood pressure that occurred by the fourth week after 5/6 nephrectomy. The antihypertensive effect of EGF was still evident at week 5. Urinary flow rate, free water clearance, and excretion of total solutes, Na+, and K+, however, were not significantly altered by EGF at weeks 2, 4, 5, or 6, suggesting a mechanism other than increased natriuresis or diuresis for this antihypertensive effect.

Acquired essential fatty acid depletion in the remnant kidney: amelioration with U-63557A.

Previous studies have demonstrated an association between renal cortical fatty acid composition and experimental models of renal injury. The present study was designed to extend these observations to the remnant kidney and to investigate the hypothesis that increased endogenous turnover of arachidonic acid metabolites results in the depletion of progenitor fatty acids. Remnant kidney cortex demonstrated a relative reduction of the essential fatty acids, linoleate and arachidonate (20 +/- 7.2% and 11 +/- 0.3%, respectively), nine weeks after subtotal nephrectomy. In addition, the monounsaturated fatty acid, oleate, was increased (48 +/- 10.6%) while its saturated progenitor, stearate, was decreased (13 +/- 4.3%). Serial evaluation of dienoic prostanoids revealed a significant increase in the renal excretion of TXB2 in rats with remnant kidneys (27 +/- 3.0, 29 +/- 1.1, and 34 +/- 3.3 ng/day vs. 21 +/- 0.8, 20 +/- 1.5, and 22 +/- 3.3 ng/day in control rats, at 3, 6, and 9 weeks, respectively). Moreover, TXB2 excretion inversely correlated with dienoic progenitor fatty acids [18:2(n-6), r2 = 0.76; 20:4(n-6), r2 = 0.79], suggesting that these events are biochemically coupled. Endogenous turnover of precursor fatty acids, confirmed by an increase in renal TXB2 excretion, preceded overt depletion of essential fatty acids by several weeks. Importantly, blockade of endogenous synthesis of TXA2 with the specific TXA2 synthase antagonist, U-63557A, restored the essential fatty acid composition to normal and ameliorated progressive glomerular destruction. Moreover, the ancillary fatty acid disturbances were attenuated by administration of U-63557A.(ABSTRACT TRUNCATED AT 250 WORDS)