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D M Brown

Publications and source records attributed to D M Brown.

At least 253 records · Page 14Linked to original sources
PubMed1984

Shwartzman reaction in streptozotocin-induced diabetic rats.

The possibility that experimental diabetes could prepare for the generalized Shwartzman reaction was investigated in female Sprague-Dawley streptozotocin-induced diabetic rats. After 48 hours, 1 week, and 9 weeks of diabetes, the rats were injected with 2 mg/kg of endotoxin, and the animals were sacrificed 2, 4, 8, and 24 hours after endotoxin. Ninety percent of the diabetic animals given endotoxin developed massive glomerular capillary fibrin deposition accompanied by marked decrease in platelet count. The age- and sex-matched nondiabetic control rats had no such changes. This marked susceptibility to endotoxin, previously only reported in pregnant rats, was present as early as 1 week of diabetes. The degree of glycemic control greatly influenced the susceptibility of diabetic rats to the generalized Shwartzman reaction. Only 28% of the diabetic animals given insulin once daily (4.6 +/- 0.3 units, mean +/- SEM) and maintaining a blood glucose level of 269 +/- 19 mg/dl developed glomerular thrombi. In contrast, the diabetic animals that did not receive insulin and had a blood glucose level of 617 +/- 21 mg/dl all developed fibrin thrombi. We conclude that the diabetic state in rats induces a unique susceptibility to the generalized Shwartzman reaction following a single injection of endotoxin, which varies with the severity of the diabetic state. Although the pathogenesis is unclear, this phenomenon may reflect abnormalities in the glomerular capillary wall and/or the coagulation system that may be important in the development of microvascular complications. Furthermore, this phenomenon may, in the animal model, mirror the increased risk of the diabetic patient to intravascular coagulation with bacterial sepsis.

Animals↗
PubMed1983

Primary structures of MCP-1 and MCP-2, natural peptide antibiotics of rabbit lung macrophages.

The microbicidal peptides, MCP-1 and MCP-2, of rabbit alveolar macrophages were purified by an improved procedure that employed preparative gel electrophoresis and high performance liquid chromatography. The peptides were arginine- and cystine-rich and lacked free sulfhydryl groups and detectable levels of carbohydrate. Complete sequence determinations revealed that MCP-1 differed from MCP-2 only by the substitution of arginine for leucine at residue 13 from the NH2 terminus and that the molecules were each single chain polypeptides of 33 amino acid residues containing three intramolecular disulfide bonds. The complete amino acid sequences of MCP-1 and MCP-2 are: (sequence in text)

Amino Acid Sequence↗
PubMed1983

Heparan sulfate--rich anionic sites in the human glomerular basement membrane. Decreased concentration in congenital nephrotic syndrome.

Recent work suggests that the normal barrier to penetration of the renal glomerular basement membrane by anionic plasma proteins may depend in part on the existence of negatively charged sites within the membrane. We describe an in vitro cytochemical method for the quantitative demonstration of anionic sites in the normal human glomerular basement membrane. In five normal subjects, ranging in age from 10 days to 57 years, the sites were distributed at regular intervals in the lamina rara externa, with a frequency of 23.8 +/- 6.8 sites per 1000-nm length of membrane. A similar distribution was observed in the basement membranes from three normal human fetuses. Ex vivo perfusion of one cadaver kidney revealed a similar distribution of anionic sites. The number of anionic sites in the glomerular basement membranes of five patients with the congenital nephrotic syndrome was reduced to 8.9 +/- 3.7 (P less than 0.001). Prior incubation of sections of normal kidney in purified heparinase resulted in a marked reduction in the number of anionic sites. We conclude that congenital nephrosis results from failure of heparan sulfate--rich anionic sites to develop in the lamina rara externa of the glomerular basement membrane.

Anions↗
PubMed1983

Hepatic and intestinal 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in genetically diabetic mice.

Cholesterol synthesis rate, as determined by 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, is characterized in the major organs of genetically diabetic mice. Both C57BL/Ks db+/db+ and C57BL/6 ob+/ob+ mice are hyperinsulinemic and insulin-resistant. These animals demonstrate loss of the circadian rhythm of hepatic reductase activity and a tendency for increased intestinal activity. As a result, proportionally more endogenous cholesterol synthesis occurs in intestinal mucosa than liver in genetically diabetic animals. Thus, the alterations in activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase which are observed in animal models of diabetes are the result of diminished insulin effect rather than insulin level.

Animals↗
PubMed1983

The effect of prenatal dexamethasone on fetal rat lung prostaglandin synthesis.

[14C]Arachidonic acid conversions were studied in homogenates of lungs from 20, 21, and 22 day fetuses with or without prenatal dexamethasone treatment. The major metabolites were in all cases 12-L-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), prostaglandin E2 (PGE2), and 12-L-hydroxy-5,8,10-heptadecatrienoic acid (HHT). Prostaglandin F2 alpha (PGF2 alpha), prostaglandin D2 (PGD2), and thromboxane B2 (TxB2) were present in small amounts. Dexamethasone treatment significantly stimulated the conversion of [14C]-arachidonic acid in fetal lung homogenates to 6-keto-PGF1 alpha, PGE2, 12-HETE, and HHT, at 20 days gestational age. This effect was dependent on the dose of dexamethasone. These results suggest that dexamethasone accelerates the maturation of the enzymes involved in prostaglandin synthesis. Because dexamethasone is also known to inhibit phospholipase A2, further studies are required to determine the overall in vivo effect of prenatal dexamethasone therapy on fetal lung prostaglandin synthesis.

Animals↗
PubMed1983

Misonidazole neurotoxicity in mice decreased by administration with pyridoxine.

A series of toxicological and pharmacological experiments was performed to test the hypothesis that alterations of pyridoxine (Vitamin B6) metabolism may play an important role in the development of misonidazole (MISO) neurotoxicity. The formation of a Schiff's base between the final reduction product of MISO, 2-amino MISO (NH2-MISO), and pyridoxal-HCl in ethanol was demonstrated. Mice receiving daily intraperitoneal injections of MISO suffered significantly less toxicity (as determined by survival, weight gain and neurological tests) when large doses of pyridoxine-HCl (PYR) were delivered concomitantly, and consequently were able to tolerate administration of more than twice as many MISO injections. PYR did not alter the pharmacokinetics of MISO, either when given simultaneously or when given by multiple repeated daily injections prior to MISO. The administration of PYR also did not alter the radiosensitization by MISO in an in vivo-in vitro cloning assay with the EMT6 tumor in BALB/c mice. If depletion or altered metabolism of pyridoxine by reduced metabolites is also responsible for the neurotoxic effects of nitroimidazoles in humans, then concomitant administration of pyridoxine (in doses greater than the molar quantity of NH2-MISO formed) should inhibit the development of such symptoms and allow administration of larger doses of MISO than are currently clinically employable.

Animals↗
PubMed1983

Glomerular manifestations of diabetes in the BB rat.

Morphologic characteristics of the renal glomeruli and tubules of BB rats with spontaneous diabetes mellitus were studied at 30 weeks' duration of diabetes. Whereas the glomerular basement membrane (GBM) was significantly thickened, no changes in the diabetic glomeruli were seen in the peripheral capillary wall area and in the fractional volumes of the mesangial cells or of the mesangial matrix. Light microscopy of the diabetic kidneys were normal, and immunofluorescent examination of diabetic glomeruli showed no increased accumulation of albumin, C3, or IgG. Diabetic rats had increased renal blood flow and glomerular filtration rates. Diabetic rats at 7, 17, and 30 weeks excreted normal amounts of urinary albumin. Thus kidneys of the BB diabetic rat differ from other experimental models of diabetes in that GBM thickening occurs in the absence of mesangial changes and of increased albuminuria. These studies suggest that the mesangium may influence glomerular permeability in diabetes, while thickening of the GBM in diabetes does not necessarily coincide with increased urinary albumin excretion. Furthermore, these results are consonant with the hypothesis that genetic factors may influence the pathological expression of diabetic nephropathy in rats.

Animals↗
PubMed1983

Arachidonic acid deficiency in streptozotocin-induced diabetes.

Fatty acid compositions of phospholipids of heart, liver, kidney, aorta, and serum from rats having streptozotocin-induced diabetes were determined and compared with those of nondiabetic controls. Linoleic and dihomo-gamma-linolenic acids were increased whereas arachidonic acid was decreased in most tissues, suggesting an impairment of delta 5-desaturase activity. Acids derived from linolenic acid were increased in some diabetic tissues from diabetic animals although the linolenic content was normal, indicating less impairment in the desaturation of the omega 3 series of fatty acids. Diabetes suppressed all polyunsaturated acids in the whole animal, but the competition between omega 3 and omega 6 acids favored the excessive suppression of long-chain omega 6 acids and an increase in the proportion of omega 3 acids in lipids of vital tissues. These changes in fatty acid composition of the phospholipids may have significant effects on cellular functions and vasoregulatory control mechanisms in diabetes.

Animals↗
PubMed1983

Tissue zinc status of genetically diabetic and streptozotocin-induced diabetic mice.

A structural and functional relationship exists between zinc and insulin. In the present study zinc concentrations of various tissues from genetically diabetic and streptozotocin-induced diabetic mice and their appropriate control mice were determined. The zinc concentrations were depressed in serum and femur of C57BL/Ks-db+/db+ mice (db/db) when compared with their nondiabetic heterozygote controls (db/m) and homozygous controls (m/m). No differences were noted in the hepatic or renal Zn concentration of the db/db, db/m, or m/m mice. Zinc supplementation in the drinking water for a 4-wk period had no effect on serum or tissue zinc concentration. Hyperzincuria was noted in the db/db mice. No differences were noted in the Zn concentration of serum or tissue in streptozotocin-induced diabetic mice compared to their controls. These data suggest that zinc deficiency may play a role in the pathogenesis of the insulin resistance present in type II (insulin independent) diabetics.

Animals↗
PubMed1983

Diabetic nephropathy. A perspective.

The earliest manifestations of clinical diabetic nephropathy, including proteinuria, hypertension, and declining GFR, represent very advanced diabetic glomerulopathy with especially prominent mesangial expansion. Mesangial expansion, by restricting glomerular capillary filtration surface and lumenal volume, stimulates compensatory mechanisms analogous to those resulting from a marked reduction in nephron number. These compensatory mechanisms involve alterations in glomerular hemodynamics designed to maintain glomerular filtration but which ultimately injure the kidney. These hemodynamic perturbations are not specific to diabetes but represent a final common pathway toward endstage renal failure that also characterizes the remnant kidney. This thesis concludes that the onset of clinical diabetic nephropathy augurs inevitable decline in kidney function, and that only studies and interventions exercised before clinical nephropathy develops can influence understanding and outcome of diabetic nephropathy.

Basement Membrane↗
PubMed1983

Glucosylated and normal human of rat albumin do not bind to renal basement membranes of diabetic and control rats.

Hyperglucosylated and normal human and rat albumin were injected intravenously into control and streptozotocin-induced diabetic rats. Binding of injected human albumin to renal basement membranes was not observed by immunofluorescence microscopy in the diabetic or control rats irrespective of the form of injected albumin. However, human albumin was found as tubular droplets in both the control and diabetic rats injected with either form of albumin. Spontaneous binding of endogenous rat albumin was observed in a linear pattern on diabetic rat glomerular basement membranes (GBM), but not in the GBM of control rats. No appreciable differences in the intensity of staining for rat albumin was observed in diabetic rats injected with either glucosylated or normal rat albumin. Similarly, no binding of rat albumin to the GBM was observed in control rats irrespective of the type of albumin injected. These studies demonstrate that binding of albumin to renal basement membranes is not dependent on glucosylation.

Animals↗
PubMed1983

Renal localization of heparan sulfate proteoglycan by immunohistochemistry.

Glomerular localization of heparan sulfate proteoglycan (HS-proteoglycan) has been studied immunohistochemically with a highly purified antiserum to bovine aorta HS-proteoglycan core protein. The specificity of the antiserum was enhanced by consecutive fibronectin and chondroitin sulfate-dermatan sulfate proteoglycan (CS-DS proteoglycan) affinity chromatography. The affinity-purified HS-proteoglycan antibody lacked cross-reactivity by enzyme-linked immunosorbent assays (ELISA) with CS-DS proteoglycan, fibronectin, laminin, and Type IV collagen. Reactivity of the antiserum with HS-proteoglycan antigen by ELISA was inhibited by HS core protein derived from CsCl density gradient centrifugation after heparinase treatment of the HS-proteoglycan. Immunofluorescent reactivity of the HS-proteoglycan antiserum was observed with bovine glomerular basement membrane, renal interstitium, Bowman's capsule, renal arterioles, and bovine aorta. No staining was seen with rat, mouse, or human glomeruli.

Animals↗
PubMed1983

Influence of heat on the intracellular uptake and radiosensitization of 2-nitroimidazole hypoxic cell sensitizers in vitro.

The effect of elevated temperature (44 degrees) on the intracellular uptake of the 2-nitroimidazole hypoxic cell radiosensitizer, misonidazole (MIS), and analogues more hydrophilic than MIS was studied in Chinese hamster ovary cells. It was found that the intracellular uptake of these compounds which enter cells by restricted passive diffusion can be enhanced approximately 4-fold when incubated at 44 degrees compared to the uptake at 37 degrees. Peak intracellular uptake (expressed as the ratio of intracellular concentration to extracellular concentration) following incubation of cells in 2 mM MIS was 100% at 44 degrees but only 25% at 37 degrees. Furthermore, a short-term nonlethal heat pulse (44 degrees for 15 min) with MIS present caused a 2-fold enhancement in uptake which was sustained for an additional 45 min at 37 degrees. This same nonlethal heat pulse was found to induce a similar enhancement in uptake even when MIS was added at subsequent time intervals at 37 degrees. The heat pulse induced a time-related enhancement of uptake at 37 degrees which increased for 1 hr and persisted for at least 6 hr. Finally, in vitro radiosensitization studies of hypoxic Chinese hamster ovary cells showed that the nonlethal heat pulse of 44 degrees for 15 min could greatly enhance the sensitization by low concentrations (0.5 mM) of MIS added after heating due to increased intracellular concentrations of the drug. MIS (0.5 mM) alone achieved a radiosensitization enhancement ratio of 1.29 (compared to irradiated hypoxic cells alone), while the addition of the short-term heat pulse, which had only a minor effect itself, achieved an enhancement ratio of 1.78.

Animals↗
PubMed1983

Factors influencing intracellular uptake and radiosensitization by 2-nitroimidazoles in vitro.

In this study it is shown that the radiosensitization of hypoxic Chinese hamster ovary (HA-1) cells in vitro by misonidazole (MIS) and other 1-substituted 2-nitroimidazoles depends on the rate and extent of intracellular uptake of these radiosensitizers, which in turn is governed by their lipophilicity [expressed as the octanol:water partition coefficient (P)]. As the lipophilicity of the compounds decreased, the rate of drug entry into the cells was slower, and below P values of approximately 0.05, peak intracellular drug concentrations were found to be lower than that of MIS (P = 0.43). In addition, the number of hydroxyl groups on the side chain of the nitroimidazole molecule influenced the uptake of drug into the cells. For compounds of similar P, but differing in the number of side-chain hydroxyl groups, the addition of a single hydroxyl group to the molecule decreased the amount of drug entering the cell by a factor of approximately 2. These compounds enter the cell by nonmediated passive diffusion since altering the energy (ATP) capacity of the cell by 2-deoxyglucose did not affect uptake. It is also shown that increases in temperature or decreases in pH can increase the intracellular uptake of MIS. For example, equal intracellular and extracellular concentrations (100% uptake) of MIS were obtained if cells were heated to 44-45 degrees C for 15 min compared to 20-40% uptake at 37 degrees C. Increases in MIS uptake by factors of 2 to 3 could be demonstrated within 30 min when cells were incubated in Hanks' balanced salt solution at pH between 6.0 and 6.3 without loss of cell viability. In addition, MIS uptake in aerobic cultured cells varied from 15 to 60% depending on the cell line and culture conditions used.

Animals↗
PubMed1982

Disturbance of pulmonary prostaglandin metabolism in fetuses of alloxan-diabetic rabbits.

[14C]Arachidonic acid conversion in lung homogenates of 28-day fetuses from control and alloxan-diabetic rabbits was studied. The major metabolites were 12-L-hydroxy-5,8,10,14-eicosatetraenoic acid and prostaglandin E2. Small amounts of 6-ketoprostaglandin F1 alpha, prostaglandin F2 alpha, and thromboxane B2 were also observed. Lung homogenates from fetuses of alloxan-diabetic rabbits convert significantly less [14C]arachidonic acid to prostaglandin E2, whereas all other metabolites were present in similar quantities compared to fetuses of non-diabetic rabbits. These studies suggest that the decreased arachidonic acid conversion to prostaglandin E2 could be partially responsible for the functional delay of lung maturation in offspring of alloxan-diabetic rabbits.

Animals↗
PubMed1982

Diabetic glomerulopathy in the uninephrectomized rat resists amelioration following islet transplantation.

Uninephrectomy is known to accelerate the development of both functional and morphological changes seen with experimental diabetic nephropathy in the rat. The present experiments utilized electron and light microscopic morphometric techniques to assess glomerular basement membrane width and the volumes of the total mesangium and its cellular and matrix components of inbred Lewis rats made diabetic at 6 weeks of age and uninephrectomized 9 days later. Immunofluorescent microscopy was used to evaluate IgG and C3 in the mesangium. The reversibility of established diabetic glomerular lesions in uninephrectomized diabetic rats after 7 months of diabetes was studied by performing intraportal transplant of neonatal pancreatic tissue. Renal biopsies were taken 2 months later in transplanted and non-transplanted animals. Islet transplantation lowered plasma glucose to normal levels (29.6 to 7.3 mmol/l) and raised plasma insulin values (6.3 to 53 muU/I). Glomerular basement membrane width in transplanted rats (268 nm) still exceeded the same measure (226 nm) in nondiabetic uninephrectomized rats. In transplanted animals volumes of the mesangium (0.51 x 10(6) micrometers 3) and of its cellular (0.27 x 10(6) micrometers 3) and matrix (0.24 x 10(6) micrometers 3) components remained higher than similar measures in control rats (0.32 x 10(6), 0.17 x 10(6) and 0.15 x 10(6) micrometers 3, respectively). Mesangial IgG in treated animals approached normal, but mesangial C3 remained similar to levels in non-transplanted diabetic control animals. These observations in uninephrectomized-diabetic rats contrast with previous observations in intact diabetic rats in which mesangial volumes and localization of immunoglobulins and complement returned to normal levels following islet transplantation.

Animals↗
PubMed1982

Tail pinch behavior and analgesia in diabetic mice.

Mild tail pinch induced "consummatory" behaviors in mice. The major tail pinch behavior appeared to be chewing with food ingestion occurring possibly as an epiphenomenon. All tail pinch behaviors were obliterated by the dopamine antagonist haloperidol; and the opiate antagonist, naltrexone, decreased eating without altering chewing. The combination of dopamine blockade and tail pinch induced jumping behavior in mice. Diabetic mice showed increased tail flick latencies to radiant heat and to the induction of tail pinch behaviors, displaying these behaviors less commonly than their homozygote and heterozygote littermate controls.

Animals↗