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V J Savin

Publications and source records attributed to V J Savin.

At least 37 records · Page 2Linked to original sources
PubMed1994

C5b-9 increases albumin permeability of isolated glomeruli in vitro.

Deposition of antibody and activation of the complement cascade are important in both naturally occurring glomerulonephritis and in experimental models including passive Heymann nephritis. We studied the effect of antibody and complement on albumin permeability of isolated glomeruli to determine the role of the terminal complement components (C5-C9) in mediating the proteinuria in nephritis. Isolated glomeruli were treated with anti-Fx1a (Heymann antibody) and then incubated them with pooled human serum, serum in which complement had been inactivated by heat, or serum deficient in C6 or C7. The albumin reflection coefficient (sigma albumin) was calculated from the volumetric response of glomeruli to transcapillary oncotic gradients produced by albumin or high molecular weight neutral dextran (252 kD). Convectional permeability to albumin (Palbumin) was calculated as 1-sigma albumin. Albumin permeability of control glomeruli was not different from 0. Albumin permeability was not altered by antibody alone but was increased to 0.65 +/- 0.04 when antibody treated glomeruli were incubated for 10 minutes with pooled serum as a source of complement. Heat treatment of serum to inactivate complement prevented the increase in permeability. Incubation for 10 minutes with serum without antibody pretreatment caused a lesser increase in permeability of isolated glomeruli (0.18 +/- 0.06). Serum deficient in either C6 or C7 did not cause an increase in albumin permeability of antibody pre-treated glomeruli, but incubation with a combination of these sera (now containing the complete cascade) increased permeability to the same extent as did pooled normal serum (0.58 +/- 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Polymorphonuclear leukocytes increase glomerular albumin permeability via hypohalous acid.

Acute glomerulonephritis is characterized by the presence of neutrophils within glomeruli and the generation of reactive oxygen species (ROS) by activated polymorphonuclear leukocytes (PMNs). Hydrogen peroxide (H2O2) and other ROS including hypothalous acids have been implicated in PMN mediated injury. To determine the role of specific ROS in PMN mediated glomerular injury, isolated rat glomeruli were incubated for 30 minutes at 37 degrees C with H2O2, with H2O2 and myeloperoxidase, or with activated PMNs. Scavengers of ROS were included in some experiments. PMNs were harvested from rat peritoneal cavity and activated with phorbol myristate acetate (PMA). Glomerular albumin permeability (Palbumin) was calculated from the volume response to an oncotic gradient. Palbumin of glomeruli incubated with H2O2 (10(-3) or 10(-1) M) was not increased, while Palbumin after incubation with H2O2 and MPO was markedly increased (0.94 +/- 0.004). Palbumin after incubation with PMA, or with non-activated PMNs was not different from that of control glomeruli, Palbumin of the glomeruli incubated with activated PMNs increased (0.85 +/- 0.01, P < 0.001). This increase in Palbumin was inhibited by superoxide dismutase, catalase, or taurine (Palbumin = 0.035 +/- 0.06, -0.39 +/- 0.10, 0.028 +/- 0.06, respectively) and ameliorated by sodium azide (Palbumin = 0.21 +/- 0.03). In contrast, dimethyl sulfoxide did not prevent the increase in Palbumin (Palbumin = 0.92 +/- 0.01). Our results show that the hypohalous acid derived from that of H2O2-MPO-halide system is capable of increasing Palbumin. We conclude that hypohalous acid may be the primary mediator of the immediate increase in glomerular protein permeability induced by PMNs.

Albumins↗
PubMed1993

Mechanisms of proteinuria in noninflammatory glomerular diseases.

Neither the initiating factors nor the proximate causes of injury that produce proteinuria in nephrotic syndrome have been clearly defined. Immune mechanisms have been postulated in minimal-change nephrotic syndrome (MCNS), focal segmental glomerular sclerosis (FSGS), and glomerular sclerosis associated with human immunodeficiency virus (HIV) infection. Circulating factors have been proposed in MCNS and FSGS, although no specific mediator has been identified. Prompt remission of proteinuria following steroid treatment and the presence of altered immune responsiveness in patients with MCNS have been used to support the participation of an immune mechanism in the pathogenesis of MCNS. Both FSGS and HIV-related nephropathy are usually steroid-resistant. Immune mechanisms are postulated in FSGS because of early recurrence after transplantation, and in HIV-related nephropathy because of the numerous associated immune abnormalities. Experimental models of nephrotic syndrome based on neutralization of glomerular charge, toxic injury to podocytes, injection of antibodies to glomerular components, or abnormalities in transgenic mice have been used to define mechanisms of glomerular injury. This review summarizes physiologic and immunologic abnormalities in MCNS, FSGS, and HIV-associated nephropathy and in several experimental models of nephrotic syndrome, and outlines the immunologic mechanisms and cellular reactions that may be responsible for glomerular dysfunction in these entities.

Animals↗
PubMed1993

Effect of superoxide exposure on albumin permeability of isolated rat glomeruli.

Reactive oxygen radicals generated by mesangial cells or by resident or infiltrating macrophages may contribute to glomerular injury in glomerulonephritis. To determine whether superoxide anions have an effect on the glomerular barrier to macromolecules, we studied the responses of isolated glomeruli after exposure to superoxide generated by xanthine and xanthine oxidase or by activated macrophages. Glomerular volumetric responses to oncotic gradients of bovine serum albumin or an impermeant neutral dextran (mol wt 252 kd0 were used to calculate the albumin reflection coefficient (sigma albumin) and convectional permeability to albumin (1-sigma albumin) of control and superoxide-treated glomeruli. Albumin permeability of control glomeruli was not different from 0 (0.02 +/- 0.01, N = 50). After 10 minutes of exposure of glomeruli to superoxide generated by xanthine oxidase, albumin permeability was increased to 0.06 +/- 0.01 (N = 50). Albumin permeability did not increase further after incubations with xanthine and xanthine oxidase for up to 60 minutes. The increase in albumin permeability was prevented by superoxide dismutase (-0.02 +/- 0.01, N = 48) but was not affected by catalase or by indomethacin pretreatment. Coincubation of glomeruli with activated macrophages also increased albumin permeability to a maximum of 0.80 +/- 0.05 (N = 17). Albumin permeability was not increased by incubation of glomeruli with phorbol myristate acetate alone or with macrophages in the absence of phorbol myristate acetate. The effect of activated macrophages on albumin permeability, like that of superoxide generated chemically, was prevented by superoxide dismutase but not by catalase or indomethacin.(ABSTRACT TRUNCATED AT 250 WORDS)

Albumins↗
PubMed1992

Vasoactive substances induce cytoskeletal changes in cultured rat glomerular epithelial cells.

Angiotensin II (ANG II), atrial natriuretic peptide III (ANP), and sodium nitroprusside (SNP) alter capillary hydraulic conductivity in isolated glomeruli. These agents also affect cyclic nucleotide levels of glomerular epithelial cells (GEC). ANG II increases cAMP, whereas ANP and SNP increase cGMP. The effects of these vasoactive substances on GEC cytoskeleton were tested by incubating cells from primary cultures or an established cell line with each agent. Changes in the cytoskeleton were assessed by staining for F-actin with Bodipy phallacidin and for tubulin with NBD-colcemid. Control cells exhibited short bundles of F-actin or stress fibers near the base of the cells. These were frequently arranged in parallel and occasionally appeared to radiate from the center of the cell. Microtubules were arranged in a fine network throughout the cell with increased density adjacent to the nucleus and within the nucleolus. Incubation of GEC with 10(-7) M ANG II, cholera toxin, or 8Br-cAMP for 2 at 37 degrees C resulted in rearrangement of F-actin into distinct stellate patterns with a decrease in the relative intensity of the peripheral staining, all concurrent with a fivefold increase in intracellular cAMP. The incubation of GEC with 10(-6) M ANP or 10(-7) M SNP for 2 h at 37 degrees C resulted in apparent disassembly of stress fibers, sparse and more diffuse fluorescence, and some increase of fluorescence at the periphery of the cells, all concurrent with a 10-fold increase in intracellular cGMP. Cytochalasin D incubation led to complete disassembly of actin filaments.(ABSTRACT TRUNCATED AT 250 WORDS)

Actin Cytoskeleton↗
PubMed1992

Measurement of albumin reflection coefficient with isolated rat glomeruli.

Macromolecular permeability of the glomerular capillary has been inferred from the clearance of endogenous protein or infused macromolecules. Permeability is increased after treatment with polycations as well as after renal injury. It has previously been shown that the capillaries of glomeruli isolated from normal mammals expand or collapse in response to transcapillary albumin gradients and that the magnitude of the changes in capillary volume and in total glomerular volume are directly proportional to the applied oncotic gradients. In the experiments presented here, the volume responses of control glomeruli and of glomeruli treated with protamine (100 to 600 micrograms/mL for up to 60 min) were used to calculate the albumin reflection coefficient, sigma albumin, and the convectional permeability, P convectional albumin = (1 - sigma albumin), of the capillary wall. Sigma albumin for normal glomeruli was about 1 (P convectional albumin = 0); sigma albumin fell to a minimum of 0.2 +/- 0.1 (P convectional albumin = 0.8 +/- 0.1) after incubation with protamine sulfate (600 micrograms/mL) for 30 min. Retraction and fusion of podocyte foot processes and denudation of the underlying matrix was seen on scanning electron micrographs of protamine-treated glomeruli. These results confirm that it is possible to study macromolecular permeability of the glomerular capillary in vitro and to calculate sigma albumin independent of hemodynamic and systemic humoral influences. This method will permit the assessment of the effects of individual mediators of glomerular injury and the study of glomeruli from kidneys affected by experimentally induced or naturally occurring renal diseases.

Albumins↗
PubMed1992

Diminished glomerular capillary hydraulic conductivity precedes morphologic changes in experimental diabetes mellitus in the rat.

Glomerular ultrafiltration coefficient, Kf, is diminished in established diabetic nephropathy. To determine whether Kf is decreased because of a decrease in capillary area, A, and or in hydraulic conductivity, Lp, glomerular Kf and morphometric parameters were measured, and Lp was calculated in glomeruli of young rats with STZ-induced DM and in control rats. STZ was administered to Fischer 344 rats that weighted 50-75 g; glomeruli were examined after 3 or 5 mo of DM, and their structure and function was compared with that of control rats. The effects of insulin or of an ACEI, enalapril, also were assessed after 3 or 5 mo. Growth of DM rats was markedly impaired, and their ratio of kidney weight to body weight was increased. Ccr was proportional to rat weight, and the ratio of Ccr to body weight was not different in DM and control rats. At 3 mo, average volume of glomeruli isolated from DM rats was less than that of glomeruli from control rats. In contrast, glomerular volume after 5 mo was equal in DM and control rats. No increase in GBM thickness or mesangial volume was observed, nor was any decrease seen in GBM area in DM rats at 5 mo. Kf was lower in DM rats than controls after 3 mo, but not after 5 mo. The Lp of DM and control glomeruli did not differ at 3 mo, but was lower in DM at 5 mo. Insulin therapy improved somatic growth and increased kidney and glomerular size in DM rats; the kidney weight/body weight ratio remained elevated.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1990

In vitro effects of cyclosporine on glomerular function.

Cyclosporine A (CsA) causes vasoconstriction and decrease in glomerular filtration rate. Experiments were conducted in isolated glomeruli to study effects of CsA of glomerular perfusion and independent of systemic or renal factors. CsA caused a dose dependent decrease in glomerular volume consistent with mesangial contraction. The ultrafiltration coefficient Kf was significantly lower after incubation in 4 X 10(-4) M CsA when compared to control (2.81 +/- 0.2 vs 5.19 +/- 0.5 nl/min X mm Hg; p less than 0.001). Hydraulic conductivity Lp was diminished by CsA from 2.69 +/- 0.11 to 1.41 +/- 0.05 microliter/min X mm Hg X cm2 (p less than 0.001). These findings demonstrate a direct effect of CsA on the glomerulus which must be considered in addition to drug-induced changes in perfusion and tubular function.

Animals↗
PubMed1990

Glomerular effects of angiotensin II require intrarenal factors.

Glomerular ultrafiltration coefficient (Kf) of glomeruli isolated from kidneys of normovolemic rats decreases following infusion of angiotensin II (ANG II). Kf from isolated glomeruli after ANG II infusion in vivo and from isolated perfused kidneys following infusion of ANG II in vitro was measured to determine whether the decrease required the presence of systemic factors. Filtration was induced in vitro and the maximum rate of change in glomerular volume was used to calculate Kf. Glomerular capillary hydraulic conductivity (Lp) was calculated from Lp = Kf/A where the basement membrane area A was calculated as 3 X pi X D2. ANG II infusion in vivo in rats diminished Lp from 3.19 +/- 0.19 to 1.96 +/- 0.13 and to 1.82 +/- 0.11 microliters.min-1.mmHg-1.cm-2, respectively. ANG II infusion into isolated kidneys caused a similar decrease in Lp (3.55 +/- 0.11 to 2.37 +/- 0.07). ANG II infusion either in vivo or during isolated kidney perfusion decreases Kf and Lp. ANG II effects do not require the presence of extrarenal factors but depend on perfusion in situ since incubation of isolated glomeruli with ANG II did not alter Kf.

Angiotensin II↗
PubMed1989

Dietary protein and glomerular response to subtotal nephrectomy in the rat.

Renal tubules and glomeruli undergo hypertrophy after partial renal ablation. Glomerular and vascular scarring and progressive renal insufficiency may later occur. In the present studies, we have documented the filtration characteristics of glomeruli isolated from rats that had undergone subtotal nephrectomy and been fed a normal protein (NP) diet (24 gm protein per 100 gm chow) or a protein-restricted (PR) diet (6 gm protein per 100 gm chow) and compared these glomeruli to those of sham-operated (SHAM) rats maintained on a normal diet. Body weight of SHAM rats increased by 30% during the 28 weeks of the study. Weight of NP rats did not change significantly, and weight of PR rats decreased by about 25%. Seven weeks after surgery, the glomeruli of NP rats were significantly larger and the ultrafiltration coefficients (Kf) were higher than those of SHAM rats; glomeruli of PR rats were of normal size but also had increased Kf. Twenty-eight weeks after surgery, the glomeruli of rats in each group were larger than at 7 weeks. Kf was no longer elevated in NP rats. Glomerular capillary hydraulic conductivity (Lp) was equal in all groups at 8 days but was increased at 7 weeks and decreased 28 weeks after surgery in NP rats. We hypothesize that glomerular hypertrophy occurs in response to renal ablation and that subsequent changes in the capillary wall lead to diminished Lp. Protein restriction in this model prevents both glomerular hypertrophy and a decrease in Lp.

Animals↗
PubMed1988

Ultrafiltration coefficients of glomeruli from human biopsies.

Ultrafiltration coefficient (Kf) was measured in vitro using glomeruli isolated from 28 human renal biopsies in order to asses both the relationship between glomerular structure and filtration characteristics and the relationship between Kf and patients' clinical state. The patients, ages 1 to 72 years, had a wide variety of renal diseases and serum creatinines of 1 to 16 mg/dl. Glomeruli were examined by light, immunofluorescence and electron microscopy, and glomerular alterations were measured. Filtration was induced in isolated glomeruli by an oncotic gradient and Kf calculated. Glomerular diameter (D), averaged for each patient, varied from 131 to 315 microns, and Kf varied from 5.7 to 51 nl/min mm Hg. Hydraulic conductivity (Lp) in 15 biopsies averaged 1.45 mu.min-1.mm Hg-1.cm-2. In order to identify the significant predictors of Kf and delineate their relationships, stepwise multiple regression analysis was performed. Kf increased with increasing glomerular size and with increasing degree of glomerular hypercellularity. D, in turn, increased with body surface area, urinary protein, and degree of capillary damage, and decreased with percent senescent glomeruli and degree of epithelial foot process broadening. Kf did not significantly correlate with clinical measures of renal function.

Biopsy↗
PubMed1986

Percutaneous transluminal angioplasty in transplant renal artery stenosis: experience and review of the literature.

Percutaneous transluminal angioplasty (PTA) was performed in five instances of renal transplant artery stenosis (RTAS) in four patients. Hypertension was present in all cases and improved after angioplasty together with reduction in medicine requirements. Abnormal renal function in four instances also improved after PTA. This reflects the current literature in which 76 of 90 patients were successfully treated by PTA (follow-up to 24 months), with two cases of recurrent stenosis, no mortality, and only a single case of graft loss. Vascular surgical repair succeeded in 130 to 180 patients, but graft loss occurred in 20 cases and recurrent stenosis in 11. Mortality was reported in five cases. Our review of the literature and experience suggests that PTA may be preferred in the treatment of RTAS.

Adult↗
PubMed1986

Filtration by superficial and deep glomeruli of normovolemic and volume-depleted rats.

We measured glomerular ultrafiltration coefficient (Kf) of isolated superficial (S) and deep (D) glomeruli of normovolemic and volume-depleted rats. Filtration was induced in vitro, and Kf was calculated from the maximum rate of change in glomerular size. Basement membrane area (A) for each glomerulus was estimated from morphometric analyses, and glomerular capillary hydraulic conductivity (Lp) was calculated by the formula Lp = Kf/A. Kf of S and D glomeruli of normovolemic rats were 2.98 +/- 0.98 and 4.25 +/- 0.07 nl . min-1 . mmHg-1, respectively. In hypovolemic rats, Kf of S glomeruli fell by approximately 50% to 1.52 +/- 0.14 nl . min-1 . mmHg-1 (P less than 0.001), whereas Kf of D glomeruli remained unchanged at 4.28 +/- 0.10 nl . min-1 . mmHg-1. Lp, calculated using the peripheral capillary area, averaged 1.98 +/- 0.09 and 1.98 +/- 0.06 microliter . min-1 . mmHg-1 . cm-2 in S and D glomeruli of normovolemic rats and 1.89 +/- 0.11 microliter . min-1 . mmHg-1 . cm-2 in D glomeruli of hypovolemic rats. Lp of S glomeruli of volume-depleted rats (0.90 +/- 0.03 microliter . min-1 . mmHg-1 . cm-2) was lower than in any of the other three samples. Mild hypovolemia causes the Kf of S glomeruli to decline, whereas Kf of D glomeruli remains constant. The decrease in Kf occurs without an alteration in capillary area and is most likely due to a decrease in Lp.

Animals↗
PubMed1986

In vitro effects of angiotensin II on glomerular function.

Effects of incubation with angiotensin II (ANG II) on isolated rat glomeruli were studied to test the hypothesis that ANG II exerts a direct effect on glomerular ultrafiltration coefficient (Kf) or hydraulic conductivity (Lp). Glomeruli of adult Munich-Wistar rats were isolated in isotonic medium and incubated at 37 degrees C in control medium or in medium containing ANG II (5.3 X 10(-9) to 5.3 X 10(-6) M) for 20 min. Glomerular volume decreased after incubation with ANG II. Relative volume decrement of unselected cortical glomeruli averaged 6 +/- 3% in 5.3 X 10(-9) or 10(-8) M ANG II and 13 +/- 3% in 5.3 X 10(-6) M ANG II; volume of superficial cortical glomeruli and deep cortical glomeruli diminished by 9 +/- 2 and 5 +/- 0.5%, respectively, after incubation with ANG II 5.3 X 10(-6) M. Glomerular oncometric response after equilibration with media of differing protein concentrations was not altered by ANG II incubation. Kf and Lp assessed in samples of glomeruli following ANG II incubation averaged 4.5 +/- 0.5 nl X mm-1 X mmHg and 2.7 +/- 0.5 microliter X mm-1 X mmHg X cm2, respectively, and were not significantly different from control values (3.8 +/- 0.3 nl X mm-1 X mmHg and 2.4 +/- 0.3 microliter X mm-1 X mmHg X cm2, respectively. In replicate studies of individual glomeruli prior to and after ANG II incubation, Kf was also not significantly altered (control Kf, 6.1 +/- 1.1; Kf after ANG II, 4.4 +/- 0.9 nl X min-1 X mmHg).(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗
PubMed1986

Filtration by rat glomeruli after expansion of extracellular fluid volume.

We studied the filtration characteristics of glomeruli isolated from the superficial and deep renal cortex of Munich-Wistar rats (180 to 300 gm) to determine whether the ultrafiltration coefficient (Kf) or hydraulic conductivity (Lp) of the glomerular capillary is altered during expansion of extracellular fluid volume with isotonic protein-free (SAL) or hyperoncotic (ALB) solutions. SAL rats were given an initial infusion of 1% dextrose in lactated Ringer's solution (0.1 ml/gm body weight) followed by a maintenance infusion for 18 to 20 hours. ALB rats were given hyperoncotic bovine serum albumin (0.01 gm/gm body weight) intravenously 2 hours before sacrifice. At sacrifice, urine flow rate and fractional excretion of sodium were increased and plasma renin activity decreased in both SAL and ALB rats. Expansion of intravascular volume was evidenced by a decrease in the hematocrit of ALB but not of SAL rats; plasma protein concentration was increased in ALB and unchanged in SAL rats. Kf and Lp were estimated during in vitro filtration. Because SAL rats were significantly larger than ALB rats, results were compared with those in separate size-matched controls. Kf of superficial glomeruli of large control and SAL rats did not differ (3.7 +/- 0.4 and 3.5 +/- 0.4 nl/min/mm Hg, respectively). Lp, of the same glomeruli, calculated as the quotient of Kf and estimated filtering area (LpD), were also comparable (1.9 +/- 0.2 and 1.8 +/- 0.2 microliter/min/mm Hg/cm2). Kf of superficial glomeruli of ALB rats was significantly greater than Kf of size-matched control rats (3.7 +/- 0.3 vs. 2.9 +/- 0.1 nl/min/mm Hg, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Effective use of streptokinase for peritoneal catheter failure.

The fibrinolytic enzyme streptokinase (streptase) was infused into the peritoneal catheter in 19 episodes of catheter failure in 16 patients. Intraabdominal bleeding prior to infusion was seen in seven of these episodes. Fibrin strands and clots were present in four additional successful cases. Streptokinase successfully relieved the obstruction in 13 episodes in 11 patients. The procedure failed in two cases of omental ingrowth and in another with catheter malposition. Streptokinase infusion also failed in two patients with Pseudomonas aeruginosa and one patient with Staphylococcus epidermidis peritonitis. Intraperitoneal streptokinase infusion is simple and free of side effects. Its use should be considered in peritoneal catheter failure, particularly in cases where bleeding or fibrin accumulation may play a role.

Adult↗
PubMed1985

Ultrafiltration coefficient of isolated glomeruli of rats aged 4 days to maturation.

In the developing rat, RBF and GFR increase in parallel. We have calculated filtration coefficient (Kf) and hydraulic conductivity (Lp) of the glomerular capillary using glomeruli isolated from Munich-Wistar rats aged 4 days to adult. Lp derived as the quotient of Kf and total basement membrane area (LpS) may represent a minimum estimate, while the quotient of Kf and the peripheral basement membrane area (LpP) may more accurately represent the value that is operative in vivo. Rats in Group 1 (age 4 to 9 days) had an average glomerular diameter of 87 +/- 3 microns, Kf of 1.7 +/- 0.3 nl/min . mm Hg and LpS and LpP, 1.9 +/- 0.2 and 4.4 +/- 0.6 microliter/min . mm Hg . cm2, respectively. Glomerular diameter and Kf were not significantly increased in rats of Group 2 (22 to 36 days), but LpS and LpP diminished to 1.5 +/- 0.1 and 2.6 +/- 0.2 microliter/min . mm Hg . cm2 because of an increase in basement membrane surface density. In older rats of Groups 3 (69 to 84 days) and 4 (about 120 days, wt 205 +/- 4 g), glomerular diameter and Kf increased progressively. Kf and diameter were greater in deep than in superficial glomeruli of Group 4. In superficial glomeruli of Group 4, LpS averaged 1.1 +/- 0.1 and LpP 2.0 +/- 0.1 microliter/min . mm Hg . cm2, and were significantly lower than the values for Group 2 but not different from those of deep glomeruli of Group 4. Relative erythrocyte ejection increased with increasing glomerular diameter, indicating decreasing capillary resistance. Our observations at consistent with the previous reports that renal perfusion, Kf, and filtration rate increase in parallel, but do not support the hypothesis that glomerular capillary hydraulic conductivity increases as the filtration barrier matures.

Age Factors↗