Immediate and long-term renal effects of fetal exposure to gentamicin

Aminoglycoside antibiotics, like gentamicin, given to pregnant females cross the placenta and accumulate in the fetal kidney, which, like the adult kidney, was found to be the major site of deposition. In young guineapigs whose mothers were given gentamicin during the week following nephrogenesis in the fetus, nephron growth was found to be retarded temporarily. In rats whose mothers were given gentamicin during the period of fetal nephrogenesis, the final number of nephrons was reduced by about 20%. In both cases, renal development was imparied. although the concentration of gentamicin in the fetal kidney was lower than that measured in the kidney of human fetuses whose mothers had received a single injection of aminoglycoside. In rats exposed to gentamicin in utero, cellular damage of the undifferentiated and differentiating renal tissues was observed. It is, there are likely that the oligonephronia observed in animals born of gentamicin-treated mothers resulted from a direct effect of the drug at early stages of nephrogenesis. When gentamicin administration to the mother was prolonged, part of the oligonephronia observed at birth might have also resulted from fetal growth retardation, secondary to adverse effects of the drug on the mother. Providing it was not associated with fetal growth retardation, the presence of high gentamicin concentrations in the fetal kidney at late stages of nephrogenesis did not affect nephron differentiation. Long-term studies of rats born with gentamicin-induced oligone-phronia showed that neither the antibiotic still present in kidney several weeks after birth, nor the injuries it caused, prevented renal growth and morphological adaptation of the nephrons to their reduced number. This also holds true for functional adaptation, except in the case of phosphate transport. Of interest is the fact that the mild oligonephronia acquired in utero after exposure to gentamicin was sufficient to cause early development of glomerular sclerosis in the adults.     

Renal function of children exposed to cyclosporin in utero.

BACKGROUND: The use of cyclosporin (CsA) has improved graft survival in transplant (Tx) patients despite its potential nephrotoxicity. Children born to transplanted women may present with intrauterine growth retardation (IUGR). On the basis of potential reduced nephron mass both in IUGR and in newborn experimental animals exposed to CsA in utero, we investigated the renal function of children >1 year of age born to women under maintenance immunosuppression, including CsA. METHODS: Fourteen children born to 12 Tx women (nine kidney, one pancreas-kidney, one heart, one liver) were investigated using inulin clearance (C(in)), para-aminohippuric acid clearance (C(PAH)), microalbuminuria, and electrolyte reabsorption rate. RESULTS: Gestational age of the 14 infants was 34+/-3 weeks and birth weight 2018+/-620 g. During pregnancy, CsA trough blood level was 234+/-115 microg/l and plasma creatinine range was 96-136 micromol/l. Two children were excluded from the study because renal investigation led to a diagnosis of hereditary nephritis (one Alport syndrome, one familial dominant focal segmental glomerulosclerosis) that was retrospectively completed in the mother. Renal function tests were finally performed in 12 children at 2.6+/-1.8 years of age: BP 94+/-7/55+/-5 mmHg, C(in) 117+/-28 ml/min/1.73 m(2), C(PAH) 545+/-124 ml/min/1.73 m(2), filtration fraction 0.23+/-0.03, microalbuminuria 4.2+/-3.5 mg/mmol. Electrolyte tubular reabsorption rates and urine concentrating capacity were normal. CONCLUSION: These results suggest that in children born to transplanted women taking CsA, renal function develops normally despite prolonged exposure in utero.     

Adverse effects of hyperglycemia on kidney development in rats: in vivo and in vitro studies.

Congenital malformations occur more frequently in the offspring of diabetic mothers. These in vivo and in vitro studies investigate the potential adverse effects of hyperglycemia on kidney development in the rat. Female rats were made hyperglycemic throughout gestation with a single injection of streptozotocin (STZ) on day 0 of gestation, or for a short period encompassing the early stage of renal organogenesis by infusing glucose from gestational days 12-16. Kidney development in the pups was assessed by determining the total number of nephrons formed in the kidney. The number of nephrons was significantly reduced (10-35%) in the pups from STZ-treated dams, as a function of hyperglycemia. There were also fewer nephrons in pups from dams given glucose infusion whose hyperglycemia was transiently higher on day 13 of gestation. The in vitro experiments were done on metanephroi removed from 14-day-old fetuses and grown for 6 days in medium containing 0, 6.9, 13.8, or 27.5 mmol/l glucose. The development of explants grown in 0, 13.8, and 27.5 mmol/l glucose was impaired compared with that of explants grown in the 6.9 mmol/l control medium, showing that the glucose concentration must be closely controlled to ensure optimum in vitro metanephros development. Thus, exposure to hyperglycemia in utero can cause a nephron deficit, which in turn may have renal consequences later in life.     

Effects of pentoxifylline, pentifylline and gamma-interferon on proliferation, differentiation, and matrix synthesis of human renal fibroblasts.

BACKGROUND: Kidneys that progress to end-stage renal failure are almost invariably characterized by the presence of tubulointerstitial fibrosis. Therapeutic interventions to halt the progressive deterioration of renal function are still limited. Pentoxifylline, pentifylline, and gamma-interferon have shown a potential benefit in the treatment of fibrotic processes in the skin and lung. Thus, the aim of the present study was the analysis of potential anti-fibrotic effects of these substances on human kidney fibroblasts in vitro. METHODS: Primary renal fibroblasts were established from human kidney biopsies and were studied in addition to two renal fibroblast cell lines. Cells were first growth arrested by withdrawal of fetal calf serum (FCS) and subsequently stimulated with 10% FCS in the presence of different concentrations of pentoxifylline (PTX), pentifylline (PTF), or gamma-interferon (IFN-gamma). Fibroblast proliferation was determined by bromodeoxyuridine incorporation and cell counts. Northern and western blot hybridizations for basic fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-beta1 were performed to analyse inhibitory effects. The effects of all three substances on matrix synthesis were evaluated by immunoblot analyses and ELISA for collagen type I and fibronectin after stimulation with TGF-beta1. Finally, differentiation into myofibroblasts was examined by double immunofluorescence staining for alpha-smooth-muscle actin and Hoechst dye H33258. RESULTS: PTX and PTF resulted in a dose- and time-dependent inhibition of proliferation in all fibroblast lines (maximum 78.9+/-6.2% at 500 microg/ml PTX). Conversely, IFN-gamma had only modest effects on fibroblast proliferation, resulting in a maximum of 36.0+/-6.1% inhibition at 500 U/ml. Northern blot hybridizations determined that FGF-2 mRNA levels in fibroblasts were decreased up to 73.7 and 91.5% by PTX (1000 microg/ml) and PTF (100 microg/ml), whereas IFN-gamma led to a reduction of 46.2% at 1000 U/ml, indicating that the inhibitory effects of all three substances may be mediated through inhibition of FGF-2 synthesis. These findings were corroborated by immunoblot analyses where again PTX and PTF had the strongest inhibitory effects. No change in TGF-beta1 mRNA levels was noted. Synthesis of cellular and secreted collagen type I was robustly inhibited by PTX and PTF, whereas IFN-gamma exerted the strongest inhibitory effect on fibronectin synthesis and secretion. In addition, IFN-gamma down-regulated the expression of alpha-smooth-muscle actin up to 73.3% (at 1000 U/ml) whereas PTX and PTF resulted in a down-regulation of up to 49.7+/-1.8 and 80.0+/-4.4% (at 1000 and 100 microg/ml) respectively. PTF was in all experiments about 10 times more potent than equimolar concentrations of PTX. CONCLUSIONS: PTX and PTF exerted robust inhibitory effects on fibroblast proliferation, extracellular matrix synthesis, and myofibroblastic differentiation. Conversely, IFN-gamma caused strong inhibition of fibronectin synthesis and alpha-smooth-muscle cell actin expression but had only weak inhibitory influences on fibroblast proliferation and collagen type I synthesis. Inhibitory effects of all three substances on proliferation may be mediated through inhibition of FGF-2 synthesis.     

The number of nephrons in the mammalian kidney: environmental influences play a determining role

Several lines of evidence, mostly derived from animal studies, indicate that changes in the fetal environment may affect the renal development. Fetal growth retardation is associated with a nephron deficit in both humans and animals. Changes in the supply of vitamin A to the fetus may be responsible for the variations in the number of nephrons in the human kidney. In utero exposure to hyperglycemia or drugs may also cause a nephron deficit.     

In utero bone marrow transplantation induces kidney allograft tolerance across a full major histocompatibility complex barrier in Swine

BACKGROUND: In utero hematopoietic stem-cell transplantation has been shown to induce donor-specific tolerance in small-animal models. However, tolerance has been difficult to achieve in large-animal studies. METHODS: Outbred swine underwent in utero transplantation of fully major histocompatibility complex (MHC)-mismatched CD3-depleted bone marrow mixed with fresh bone marrow to achieve a final CD3 content of 1.5%. Transplantation was performed at 50 to 55 days' gestation and two animals survived long term and demonstrated multilineage peripheral blood hematopoietic chimerism. These two long-term survivors were analyzed for in vitro evidence of donor-specific tolerance by mixed leukocyte reaction (MLR), cell-mediated lysis (CML), and antibody testing and in vivo by kidney transplantation. RESULTS: Both animals demonstrated in vitro donor-specific unresponsiveness by MLR and CML and did not demonstrate anti-donor antibody production. Donor matched kidney transplants were performed without immunosuppression and functioned for more than 100 days, with no evidence for rejection. CONCLUSIONS: The authors demonstrate conclusively that in utero transplantation of fully MHC-mismatched bone marrow in swine can lead to engraftment and stable multilineage hematopoietic chimerism and tolerance to postnatal donor MHC-matched kidney transplantation without the need for immunosuppression.     

The Effect of Serum ALG Concentrations on Results Following Renal Transplantation

Seventy-three recipients of renal allografts from cadaver donors, and 121 recipients of kidneys from living related donors were studied to determine whether there were any differences in posttransplant results between patients that had a high average serum concentration of ALG (>/=800 microg/ml) during the two weeks of ALG therapy and patients that had low serum levels (/=800 microg/ml when high risk patients with diabetes mellitus were excluded. There were significantly also fewer rejection episodes at three months in recipients of living related kidney grafts that had serum ALG levels >/=800 microg/ml. When high risk diabetics or patients older than 40 were excluded from the related recipients, the number of rejection episodes was still significantly less in patients with high serum ALG levels. There was significantly less kidney loss 24 and more months posttransplant in recipients of kidneys from living related donors whether or not high risk patients were excluded. These results support previous reports from the University of Minnesota indicating ALG is a safe and effective immunosuppressive agent in renal allograft recipients.     

Aminopenicillins in urology.

The use of ampicillin and amoxicillin remains a mainstay of urologic prophylaxis and therapy, however ampicillin's effectiveness against aerobic gram-negative bacilli, especially Escherichia coli has steadily decreased over time due to increasing beta-lactamase-induced resistance. Presently, up to 40-60 percent of E. coli may be ampicillin resistant. Since there have been no major resistance problems with enterococci, ampicillin remains the preferred anti-enterococcal antibiotic in penicillin-tolerant patients. Ampicillin/beta-lactamase inhibitor combinations are preferred when penicillins are used to treat ampicillin resistant aerobic gram-negative bacillary urinary tract infections (UTIs). One response to the problem of resistance has been short-course or single-dose therapy for uncomplicated cystitis in immunocompetent adults, which is less likely to cause side effects, bacterial resistance, or alterations in bowel flora. Another response has been to combine the aminopenicillins with beta-lactamase inhibitors such as clavulanate or sulbactam.     

Long-term effects of in utero exposure to cyclosporin A on renal function in the rabbit

The number of pregnant women who receive cyclosporin A (CsA) after transplantation or for autoimmune disease has increased. CsA and its metabolites can cross the placental barrier and thus interfere with fetal development. It was shown previously that rabbits that were exposed in utero to 10 mg/kg per d CsA from the 14th to the 18th day of gestation presented a 25% nephron reduction. Thus, this study was conducted to assess the long-term systemic and renal effects of a CsA-induced nephron reduction. Twenty-two pregnant New Zealand white rabbits were randomly divided into two groups: Twelve received 10 mg/kg per d CsA from day 14 to day 18 of gestation, and 10 were used as controls. Rabbits that were born to these animals were evaluated at 4, 11, 18, and 35 wk of life. Pups that were exposed antenatally to CsA presented first a permanent nephron deficit; second, glomerular, tubular, and intrarenal hemodynamics dysfunction; third, enlarged kidneys with numerous tubular and glomerular lesions; and, fourth, an endothelin-dependent systemic hypertension that worsened with age. In utero exposure to CsA induced a nephron reduction that led to systemic hypertension and progressive chronic renal insufficiency in adulthood. A long-term clinical survey is mandatory in infants who are born to mothers who were treated with cyclosporin during pregnancy.     

Pentoxifylline reduces in vitro renal myofibroblast proliferation and collagen secretion

Interstitial myofibroblasts (MF) are cells with features of both smooth muscle cells and fibroblasts. They have been universally recognized in situations of tubulointerstitial injury, where their presence has been shown to be a marker of disease progression. The objective of this study was to determine if functions of MF relevant to fibrogenesis can be modified in vitro by the phosphodiesterase inhibitor pentoxifylline (PTX). MF were obtained from sub-culture of normal rat kidney explant outgrowths maintained in DMEM + 20% fetal calf serum (FCS), supplemented with antibiotics. Cells were characterized on the basis of growth characteristics and immunohistochemistry. MF constituted >95% of cells at passage 3. Cell culture media was supplemented with the potential antagonist PTX alone (0, 1, 10, 100 microg/ml) and in combination with TGFbeta(1) (5 ng/ml). Population kinetics, proliferation and collagen production were determined from cell growth, [(3)H]thymidine incorporation and [(3)H]proline incorporation in collagenous proteins, respectively. Both serum-stimulated population growth and proliferation were reduced in a linear fashion by 1, 10 and 100 microg/ml PTX (all p < 0.05 versus 0 microg/ml). Effect of PTX on cell population growth was however reversible when PTX was removed. Basal collagen secretion was decreased by PTX at 10 and 100 microg/ml (p < 0.05 versus 0 microg/ml although layer collagen remained unchanged. Collagen production (secreted and cell layer) was augmented by 5 ng/ml TGFbeta(1). These effects on collagen production were partially reduced when 100 microg/ml PTX was added. The authors conclude that myofibroblast function can be altered with agonists/antagonists. Attempts to down-regulate fibrogenic functions of MF may therefore offer a valuable therapeutic strategy.     

Compensatory renal growth after unilateral nephrectomy in the ovine fetus.

Unilateral nephrectomy of the adult animal results in compensatory renal growth but does not involve formation of new nephrons. It is not clear whether compensatory growth can occur during the period of active nephrogenesis in utero and if so, whether more nephrons can be formed. Male ovine fetuses (n = 20) underwent unilateral nephrectomy (n = 10) or sham nephrectomy (n = 10) at 100 d of gestation (term, 150 d). After 27 to 34 d, ewes and fetuses were killed and the right kidney of each fetus was removed and weighed. The wet weight of the right kidney was greater in the unilaterally nephrectomized fetuses (16.3 +/- 1.3 g compared with 12.2 +/- 0.7 g; mean +/- SEM, P < 0.05) as was the kidney to body weight ratio (5.2 +/- 0.3 g/kg compared with 3.8 +/- 0.2 g/kg; P < 0.001). Nephron number in the right kidney was estimated by an unbiased stereologic technique. There was a 45% increase in the number of nephrons in the kidneys from unilaterally nephrectomized animals compared with the kidneys from sham-operated animals (530,763 +/- 37,136 nephrons in the unilaterally nephrectomized group compared with 365,672 +/- 36,016 nephrons in the sham-operated group; P < 0.01). Mean glomerular volume was lower in the unilaterally nephrectomized group; however, total glomerular volume per kidney was not different between groups. This study demonstrates that there is a significant amount of compensatory growth and nephron endowment in a remaining kidney after unilateral nephrectomy during the period of active nephrogenesis in the sheep. This is the first time such events have been shown to occur in utero.     

Urinary cotinine as an objective measure of cigarette smoking in chronic kidney disease.

BACKGROUND: Smoking is a modifiable behaviour that may hasten the progression of chronic kidney disease (CKD). Cotinine, a nicotine metabolite, is measurable in body fluids, including urine, and can be utilized as an objective measure of smoking exposure. Its use has not been examined in the CKD population. METHODS: In this cross-sectional study, we evaluated use of 24-h urinary cotinine excretion (Ucot) as a quantitative index of smoking exposure in a CKD population. Methods of comparison included self-report and expired air carbon monoxide (eCO) as standard measures of smoking exposure. Assessments of kidney function included estimated glomerular filtration rate (eGFR) and 24-h urinary protein (Uprot) excretion. RESULTS: Sixty-one patients were enrolled, of whom 12 were excluded for incomplete urine collections. Of the remaining, 77% were active current smokers (mean cigarettes smoked: 12+/-7 per day). The mean eGFR was 47+/-25 ml/min/1.73 m2 with no significant differences among non-smokers. The mean eCO and Ucot were significantly higher in smokers vs non-smokers (12.5+/-6.9 ppm and 1.3+/-1.1 ppm and 1685.87+/-922.77 microg/d and 134.18+/-445.03 microg/d, respectively, P<0.001 for both). Ucot was weakly correlated with eGFR (R=0.40, P=0.005), but not with Uprot (R=0.09, P=0.54). In multivariate analyses, daily cigarette consumption and eCO were the only significant predictors of Ucot (P<0.05 for both). CONCLUSION: In this CKD cohort, Ucot is correlated with commonly used measures of smoking exposure and is minimally influenced by underlying renal function, demonstrating its potential utility in clinical trials examining change in smoking behaviour and effects on renal injury.     

JB3, an IGF-I receptor antagonist, inhibits early renal growth in diabetic and uninephrectomized rats

Biochemical evidence suggests that insulin-like growth factor I (IGF-I) may play an important role as a mediator of kidney growth. In the present study, an IGF-I receptor antagonist (JB3) was synthesized, and its effect on the renal growth that follows the induction of diabetes or unilateral nephrectomy (UNx) was examined. JB3 was generated by solid phase peptide synthesis. Its activity as an IGF-I antagonist was confirmed in an opossum kidney cell line from its inhibitory effect on the increase in thymidine incorporation into DNA induced by recombinant human IGF-I. Male Wistar rats were anesthetized with halothane and subjected to either the induction of diabetes by streptozotocin (intravenous 60 mg/kg) for 4 d (control animals received citrate buffer) or UNx for 11 d (control animals were sham operated). JB3 was delivered by subcutaneous infusion using an osmotic minipump implanted 3 d before the induction of diabetes or UNx. Kidney wet weight, DNA, and protein all were significantly higher 4 d after the induction of diabetes (24%) or 11 d after UNx (55%). Dose-response studies (1 to 30 microg/kg per day) showed JB3 administration to inhibit the increase in kidney growth in both diabetic and UNx rats. The increase in kidney wet weight, DNA, and protein was significantly lower in UNx rats that were treated with JB3 10 microg/kg per day (P: < 0.05) than in saline vehicle controls but was abolished in diabetic rats that were treated with JB3 3 microg/kg per day (P: < 0. 01). Increasing the dose of JB3 to 30 microg/kg per day was associated with a decrease in its inhibitory effect, resulting in bell-shaped dose-response curves. JB3 administration had no effect on the blood glucose concentration or food consumption by either diabetic or nondiabetic animals. The results support the concept of IGF-I as an important mediator of the early renal growth that follows the induction of diabetes or UNx in the rat.     

Midkine is involved in kidney development and in its regulation by retinoids

In the kidney, in which development depends on epithelial-mesenchymal interactions, it has been shown that retinoids modulate nephrogenesis in a dose-dependent manner in vivo and in vitro. Midkine (MK) is a retinoic acid responsive gene for a heparin-binding growth factor. The aim of the present study was therefore to quantify the expression of MK mRNA during renal development in the rat, to analyze the regulation of MK expression by retinoids in vivo and in vitro, and, finally, to study the role of MK in rat metanephric organ cultures. The spatiotemporal expression of MK in fetal kidney was studied. In control rats, MK expression is ubiquitous at gestational day 14, i.e., at the onset of nephrogenesis. On day 16, MK is expressed in the condensed mesenchyme and in early epithelialized mesenchymal derivatives. On gestational day 21, MK is rather localized in the nonmature glomeruli of the renal cortex. In utero exposure to vitamin A deficiency did not modify the specific spatial and temporal expression pattern of MK gene in the metanephros, although a decrease in mRNA expression occurred. In metanephroi explanted from 14-d-old fetuses and cultured in a defined medium, expression of MK mRNA was found to be stimulated when retinoic acid (100 nM) was added in the culture medium. Finally, in vitro nephrogenesis was strongly inhibited in the presence of neutralizing antibodies for MK: the number of nephrons formed in vitro was reduced by approximately 50% without changes in ureteric bud branching morphogenesis. These results indicated that MK is implicated in the regulation of kidney development by retinoids. These results also suggested that MK plays an important role in the molecular cascade of the epithelial conversion of the metanephric blastema.     

Stereological assessment of renal development in a baboon model of preterm birth

At the time when most preterm babies are delivered, nephrogenesis is still ongoing, with the majority of nephrons normally formed during the third trimester of pregnancy. The extrauterine environment, however, is suboptimal for organogenesis, and therefore renal development is likely to be adversely affected by preterm birth. In the long-term, there is emerging evidence of high blood pressure and renal dysfunction amongst young adults born preterm. There is little knowledge to date, however, regarding the effects of preterm birth on renal structural development, perhaps due to the lack of an appropriate animal model. We have demonstrated that the baboon (Papio sp.) has a similar time course of nephrogenesis as the human kidney, and the baboon neonate can also be cared for in the same manner as a human neonate following preterm birth. Through a series of studies assessing renal development in the baboon model of preterm birth, involving the use of gold-standard stereological techniques, we have demonstrated that nephron endowment in the preterm baboon kidney is not reduced. Furthermore, antenatal glucocorticoid exposure prior to preterm delivery was associated with an increase in mature nephrons. There was, however, evidence of morphological abnormalities in a variable percentage of the glomeruli formed ex utero. Further research is therefore essential in order to establish what factors are involved in contributing to the glomerular abnormalities, and to identify ways in which 'normal' renal development can be conserved and optimised in the extrauterine setting.     

Case of lead nephropathy due to chronic occupational lead exposure

We report a case of a patient with chronic kidney disease likely due to lead nephropathy. He was a manufacturer of Buddhist altar fittings and had chronic lead exposure. The blood lead level was 41 microg/dL and urinary lead excretion at 24 hours after the administration of ethylenediaminetetraacetic acid (EDTA)was 600 microg (first time)and 687 microg (second time), respectively. Urinary lead excretion at 72 hours was 834 microg (first time) and 1,071 microg (second time), respectively. Renal biopsy showed interstitial fibrosis and focal monocyte infiltration. Lead content in the renal biopsy specimen was 130 ng/g of wet weight. We preformed weekly EDTA chelation therapy twelve times. During the therapy, serum creatinine was 1.1 mg/dL. The chelation therapy was interrupted by an episode of acute renal failure due to hypotension and heart failure. Urinary lead excretion exceeding 600 microg at 72 hours after chelation therapy indicated a lead body burden capable of causing lead nephropathy. In this case, urinary lead excretion exceeded 600 microg at 72 hours. Based on the report that repeated lead chelation therapy can slow the progression of non-diabetic chronic kidney disease with 72-hour-urinary lead excretion of 60-600 microg, we performed chelation therapy. This case suggests that lead nephropathy currently can occur in Japan. It is possible that renal dysfunction from lead nephropathy is reversed by minimizing lead exposure and chelation therapy. Lead nephropathy should be included in the differenitial diagnosis of causes of chronic kidney disease and occupational and environmental lead exposure should be investigated carefully during the medical history.     

Distribution of renal medullary hyaluronan in lean and obese rabbits

BACKGROUND: Obese individuals have an expanded interstitium in the renal inner medulla (IM), which stains positively with periodic acid-Schiff and Alcian blue. In obese dogs, the IM is also expanded, with hyaluronan (HA) content being 2.4 times control. METHODS: We determined the anatomic pattern of renal HA deposition following weight gain, using an animal model of obesity consisting of young rabbits (N = 10), representing animals entering into the study, lean rabbits (N = 19), fed a control diet, and obese rabbits (N = 19), fed a high-fat diet (15% fat, by fortifying with corn oil and lard, in a ratio of 2:1) for two to three months. Tissue was papain digested, and HA was recovered in a phosphate or a Tris buffer and detected by an indirect immunoabsorbent competition assay. RESULTS: Rabbits fed a high-fat diet for 8 to 12 weeks gained weight (37%) and became mildly hypertensive (10 mm Hg). In lean rabbits, HA was low in the renal cortex (6 +/- 30 microg/g tissue), increased steadily across the outer medulla (OM; 79 +/- 28 microg/g tissue) and was uniformly high in the IM (192 +/- 28 microg/g tissue) when recovered in a Tris buffer; these levels of tissue HA did not change during the three-month period of dietary intervention. In obese rabbits, the renal medullary interstitium was expanded and stained intensely with periodic acid Schiff and Alcian blue, and tissue HA was elevated in the IM (448 +/- 25 microg/g tissue) but not the cortex (5 +/- 25 microg/g tissue) or the OM (85 +/- 25 microg/g tissue). The significant difference was due to those IM samples taken from the renal papilla; IM samples from the body of the kidney did not significantly differ among the lean, obese, and young rabbits. CONCLUSION: The elevated renal HA associated with weight gain is limited to the IM and occurs most consistently in the papilla, which is the region of the kidney that is most vulnerable to distention caused by elevated renal interstitial hydrostatic pressure.     

Influence of long-term diabetes on renal glycogen metabolism in the rat

BACKGROUND/AIMS: The effects of acute insulin deficiency on the kidney have been investigated in animal models of experimental diabetes; however, the impact of long-term diabetes has not been determined. METHODS: We measured renal glycogen contents in streptozotocin (STZ)-diabetic rats 3 weeks (n = 12) or 9 months (n = 12) after the induction of diabetes, and in 2 groups of control rats of similar age (n = 16 and n = 12, respectively), in the fed state and after a 24-hour fast. RESULTS: Diabetic rats had high glucose levels, low insulin but normal glucagon concentrations in portal blood. In the fasting state, kidney glycogen content was very low in both young control and young diabetic rats (54 +/- 15 and 189 +/- 26 microg/g, respectively, mean +/- SD); in contrast, glycogen levels were markedly elevated in rats with long-standing diabetes as compared to old nondiabetic animals (2,628 +/- 1,023 +/- and 1,968 +/- 989 microg/g of diabetic rat, fasting and fed, respectively, p < 0.001 vs. 0 +/- 0 and 4 +/- 6 microg/g of control rats). On electron microscopy, large glycogen clusters were localized to the renal tubules. Kidney phosphorylase activity was higher, and synthase activity lower in diabetic than control rats (p < 0.05 for both), whereas kidney glycogen was strongly related to plasma glucose levels, suggesting that the enzyme changes were secondary to glycogen accumulation itself. Renal hexosephosphates and fructose-2,6-bisphosphate contents were both increased in long-term diabetic rats (p < 0.05), implying enhanced fluxes through both glycolysis and gluconeogenesis. CONCLUSION: In chronic, untreated diabetes glycogen accumulates in the renal tubules; prolonged hyperglycemia is the sole driving force for this phenomenon.     

Evaluation of decapsulation of the canine kidney on renal function following acute ischemia.

Previous reports have suggested that the ischemic kidney develops a compression syndrome due to intracapsular edema and that decapsulation may improve function. This report evaluates renal function in ischemic kidneys with and without decapsulation. Thirteen dogs underwent creation of a split urinary bladder and the formation of bilateral cystocutaneous fistulas for measurement of split renal function. After systemic heparinization the suprarenal aorta was crossclamped for $\text{1}\text{1}\text{2}$ hr. One kidney underwent removal of the anterior aspect of the capsule and the other intact kidney served as the control. The animals received 1000 ml/24 hr, 0.15 N NaCl solution IV postoperatively. Urine volume, osmolarity, creatinine, and sodium were quantitated during three consecutive 24-hr periods. Seventy-two hours following the ischemic injury, the dogs were reexplored. Renal interstitial pressures were measured and renal biopsies and an intravenous pyelogram performed prior to sacrifice. Significantly greater urine volume was produced by the decapsulated kidney (302 ± 35 ml/24 hr) than by the nondecapsulated kidney (173 ± 39 ml/24 hr). Urine sodium excretion/24 hr was consistently greater from the decapsulated kidney than from the intact kidney while urine osmolarity at 72 hr from the decapsulated kidney was 584 ± 76 and from the intact kidney was 325 ± 76 mosmole/liter (P < 0.05). Renal interstitial pressures were significantly greater in the intact kidney than in the decapsulated kidney. These data suggest that release of intercapsular compression can improve renal function in the ischemic canine kidney.     

Beneficial effects of perfusion of atrial natriuretic factor in acute post-ischemic renal insufficiency in the rat]

Owing to its capacity of increasing glomerular filtration, potential beneficial effects of atrial natriuretic factor (ANF) were assessed during a post ischemic acute renal insufficiency in rats. Renal insufficiency was obtained by clamping the renal artery during 30 min., and by performing a reperfusion during 2 hours in uninephrectomized rats. Three groups were defined: a control group where animals were submitted to an operation procedure without renal artery clamping, a control group were animals received a physiological serum perfusion (1.5 ml/h) during the renal reperfusion time and an experimental group where animals were administrated a rat 1-28 ANF perfusion (5 microns/ml in NaCl 0.9%, 1.5 ml/hour) during the reperfusion time. Insulin clearance (1.0 + 0.05 ml/h vs 0.7 + 0.05 ml/h, p < 0.01), and diuresis (32.9 +/- 3.6 microliters/min. vs 7.5 +/- 0.23 microliters/min., p < 0.01) were significantly higher in rats which were administrated a NaCl 0.9% perfusion. Histologically, a significant decrease in kidney weight and in he percentage of diseases nephrons was observed after reperfusion in ANF treated rats. The results obtained demonstrate that ANF perfusion in case of post ischemic acute renal insufficiency in rats improves the recovery of renal function and reduces the renal histologic lesions.