Search PubMed⌕ Search

Biomedical subjects

J D Williams

Publications and source records attributed to J D Williams.

At least 109 records · Page 6Linked to original sources
PubMed1995

Elevated D-glucose concentrations modulate TGF-beta 1 synthesis by human cultured renal proximal tubular cells. The permissive role of platelet-derived growth factor.

Interstitial fibrosis is a marker of progression of renal impairment in diabetic nephropathy. Transforming growth factor (TGF)-beta 1 is one of a group of pro-fibrotic cytokines and growth factors that have been associated with the development of interstitial fibrosis. We have examined the modulating influence of glucose on the production of TGF-beta 1 by cultured human proximal tubular cells. Incubation of growth-arrested human proximal tubular cells (HPTC) (72 hours in serum free medium) in 25 mmol/L D-glucose resulted in increased expression of TGF-beta 1 mRNA (as assessed by reverse transcription polymerase chain reaction). This was apparent after 6 hours and increased up to 120 hours exposure. TGF-beta 1 secretion, however, as measured by specific enzyme-linked immunoassay, was unaffected by exposure to 25 mmol/L D-glucose. Sequential stimulation of HPTC, first with 25 mmol/L D-glucose for 48 hours and then with platelet-derived growth factor (PDGF) isoforms, resulted in a dose-dependent secretion of TGF-beta 1. Pre-exposure to 5 mmol/L D-glucose or 25 mmol/L L-glucose did not prime for TGF-beta 1 release. At 50 ng/ml PDGF this effect was greatest for the AA isoform (AA 31.4 +/- 7.1, AB 20.98 +/- 8.9, BB 7.8 +/- 2.2, P < 0.05 for all versus control, n = 3, mean +/- SEM ng/10(6) cells/24 hours). These effects were blocked by the addition of antibody to the PDGF alpha-receptor. TGF-beta 1 secretion was inhibited in a dose-dependent manner by pretreatment with cyclohexamide, but was not affected by pretreatment with actinomycin D. Stimulation of HPTC with a single dose of PDGF induced TGF-beta 1 mRNA; however, only after application of a second dose of PDGF (after TGF-beta 1 mRNA induction) did TGF-beta 1 protein secretion occur. We also demonstrated that PDGF stimulation of HPTC induced an inherently more stable TGF-beta 1 mRNA transcript. These findings demonstrate that elevated D-glucose concentration alone is insufficient to lead to increased TGF-beta 1 secretion by HPTC despite increased mRNA expression. However, application of a second stimulus such as PDGF, when TGF-beta 1 mRNA expression is increased, leads to increased protein synthesis and secretion of TGF-beta 1. This implies that elevated glucose concentrations might prime proximal tubular cells for TGF-beta 1 synthesis and thus contribute to the development of interstitial fibrosis.

Base Sequence↗
PubMed1995

Source of peritoneal proteoglycans. Human peritoneal mesothelial cells synthesize and secrete mainly small dermatan sulfate proteoglycans.

This study describes experiments that compare the proteoglycans (PGs) extracted from the dialysate from patients receiving continuous peritoneal ambulatory dialysis (CAPD) with those secreted by metabolically labeled human peritoneal mesothelial cells in vitro. The PGs isolated from both sources were predominantly small chondroitin sulfate/dermatan sulfate PGs. Western blot of the core proteins obtained after chondroitin ABC lyase treatment with specific antibodies identified decorin and biglycan. With [35S]sulfate and [35S]methionine as labeling precursors it was shown that dermatan sulfate rather than chondroitin sulfate were the major glycosaminoglycan chains and that decorin was the predominant species. These data provide the first evidence that human peritoneal mesothelial cells may be the principal source of PGs in the peritoneum. Given the proposed functions of decorin and biglycan, the results suggest that these PGs may be involved in the control of transforming growth factor-beta activity and collagen fibril formation in the peritoneum.

Ascitic Fluid↗
PubMed1995

Clinical parameters and dietary intake of midwestern adolescent females.

The purpose of this study was to investigate the correlation between assessed clinical parameters and dietary intake of adolescent females in the midwest of the United States. A total of 60 subjects participated in the study over a period of four months. Several laboratory tests along with 24-hour dietary recall were statistically analyzed. Results showed that one third of the outpatient group had serum albumin below normal range and were of marginal health status. Also, several parameters (wt., MAC, TSF, MAMA) correlated with anthoropometric measurements. Other factors: Hgb, Hct, and TLC correlated with protein intake.

Adolescent↗
PubMed1995

P1 blood group phenotype, secretor status in patients with urinary tract infections.

We have examined the distribution of P antigen, Lewis blood group phenotypes and secretor status of 65 patients with E. coli UTI (20 asymptomatic bacteriuria, 20 cystitis and normal radiology, 25 reflux nephropathy) and 45 controls who have never experienced a UTI episode. The distribution of Lewis blood group antigens was similar in all UTI groups and in the controls. The incidence of non-secretors in the reflux nephropathy group was similar to that in controls (24% versus 31%). The P1 phenotype was present in 100% of patients with asymptomatic bacteriuria, 80% with cystitis and controls and only 44% with reflux nephropathy. The combined P1/non-secretor phenotype was observed in 45% of patients with asymptomatic bacteriuria, 30% with cystitis, 12% with reflux nephropathy and in 22% of control healthy individuals. P2/secretor phenotype was demonstrated in 44% of patients with reflux nephropathy and in only 11% of controls. Our data suggest that having P2 blood group protects against asymptomatic colonization of the urinary tract, but is associated with the type of infection responsible for scarring in reflux nephropathy. It also appears that being a non-secretor does not predispose to renal scarring and that combined P2/secretor phenotype may be linked with susceptibility to reflux nephropathy.

ABO Blood-Group System↗
PubMed1994

Human neutrophils are selectively activated by independent ligation of the subunits of the CD11b/CD18 integrin.

The yeast cell wall preparation zymosan is a particulate stimulus for human neutrophils (PMNs). Unopsonised zymosan particles bind to the PMN CD11b/CD18 integrin and are phagocytosed, leading to activation of the 5-lipoxygenase pathway and release of the lipid chemotaxin leukotriene B4 (LTB4). Specific monoclonal antibodies (mAbs) to CD11b and to CD18 were used in the present study to evaluate the contribution of each chain to these processes. All four anti-CD18 mAbs but none of five anti-CD11b mAbs dose-dependently blocked PMN phagocytosis of zymosan. Nevertheless, all anti-CD11b mAbs and all anti-CD18 mAbs significantly inhibited zymosan-stimulated LTB4 release in a dose-dependent manner. In addition, there was a dose-dependent stimulation of LTB4 release resulting from the specific ligation and cross-linking of either chain of the integrin heterodimer. Thus zymosan-stimulated LTB4 release is initiated by signals from both chains of the CD11b/CD18 integrin, whereas only CD18 is essential for phagocytosis.

Antibodies, Monoclonal↗
PubMed1994

Heterophils are decisive components in the early responses of chickens to Salmonella enteritidis infections.

Chickens injected with 5-fluorouracil (5-FU) were found to have a three- to nine-fold reduction in circulating heterophils without a reduction in the number of circulating mononuclear cells. A 50% organ invasive dose (ID50) for orally and intravenously administered Salmonella enteritidis (SE) was established in the heteropenic chickens. When challenged orally, about 150-fold fewer SE cells were required for organ invasion in the 5-FU-treated chickens; whereas an intravenous challenge of the heteropenic chickens required about 4000-fold fewer bacteria for organ invasion than found in the control birds. Significant (P < 0.0001) SE-dose-dependent reductions in body weight with increased mortality and lesions in the eye, heart, and thymus were found in the 5-FU-treated birds versus the control birds. Alternatively, the control birds had significantly more intestinal lesions than the 5-FU-treated birds. No consistent significant hepatic lesions were observed in either treatment group. Under the conditions used in these experiments, 5-FU treatment of chickens caused otherwise subclinical SE infections to rapidly become clinical infections with more severe extraintestinal organ infections, whereas the control chickens had infections that were primarily restricted to the intestine. These findings suggest that the heterophil is extremely important in controlling both initial SE organ invasion and subsequent disease pathogenesis in chickens.

Animals↗
PubMed1994

The effect of methylprednisolone on monocyte eicosanoid production in patients with multiple sclerosis.

The in vitro effect of methylprednisolone on prostaglandin E2 (PGE2), leukotriene B4 (LTB4) and thromboxane B2 (TXB2) synthesis by adherent monocytes was examined using samples of peripheral blood from 15 patients with multiple sclerosis and 18 normal controls. Eicosanoid production by monocytes was reduced in patients compared with controls and there was a dose-dependent inhibitory effect of methylprednisolone on eicosanoid production in both groups. In vitro production of PGE2 and TXB2 but not LTB4 was reduced in patients with multiple sclerosis following intravenous treatment with methylprednisolone compared with pretreatment samples. In a separate cohort of 20 patients with multiple sclerosis and 15 controls, the in vitro inhibition of PGE2 release by methylprednisolone was not associated with reduced pokeweed-mitogen-stimulated immunoglobulin G synthesis by peripheral blood mononuclear cells. These results suggest that methylprednisolone inhibits monocyte-macrophage function, but this effect is not specific to patients with multiple sclerosis.

Adult↗
PubMed1994

Heritabilities for antler characteristics and body weight in yearling white-tailed deer.

Heritabilities for two body weights and five antler characteristics were estimated for a captive white-tailed deer (Odocoileus virginianus) herd maintained by the Texas Parks and Wildlife Department. Single male breeding pens with 10-14 female deer were used for five consecutive generations. To minimize selection and maintain a broad genetic base, different sets of sires and as many different dams as possible were randomly assigned as breeders each generation. All deer were accurately predigreed by sire and dam and, except for birth weight, traits were measured at 1.5 years of age. Heritabilities were estimated utilizing (1) sire and within-sire components of variance, and (2) regression of male progeny performance on sire performance. Theoretically, these procedures estimate the amount of additive genetic variance present in a population without indication of non-additive genetic (dominance and epistasis) and maternal effects. Heritabilities ranged from 0.00-0.17 (birth weight), 0.58-0.64 (body weight), 0.22-0.56 (antler points), 0.47-0.70 (main beam length), 0.03-0.43 (antler spread), 0.80-0.89 (basal circumference) and 0.71-0.86 (antler weight). These heritabilities, except for birth weight, suggest that substantial genetic change could be expected from individual selection if realistic selection differentials were used.

Animals↗
PubMed1994

Relative roles of elastase and reactive oxygen species in the degradation of human glomerular basement membrane by intact human neutrophils.

Glomerular basement membrane (GBM) damage and proteinuria occurring during the early phase of acute glomerulonephritis are often neutrophil (PMN) dependent. The present study sought to identify the potential roles of PMN derived elastase and reactive oxygen species (ROS) in the pathogenesis of glomerular basement damage in an homologous in vitro model of anti-GBM nephritis using intact PMN. Human PMN (5 x 10(6)), incubated with human GBM (0.5 mg) pretreated with human anti-GBM IgG, degraded 10.3 +/- 1.1% of the GBM type IV collagen in six hours (8 micrograms/hr), and underwent a two-hour respiratory burst. The same number of sonically disrupted PMN solubulized 22.4 +/- 5.1% of GBM under the same incubation conditions. The inclusion of the elastase inhibitors alpha 1 proteinase inhibitor (alpha 1Pi), and a smaller highly-specific synthetic compound (L658,758), reduced degradation by PMN homogenates by 84.8% and 85.7%, respectively, whereas they were only able to inhibit intact PMN mediated degradation by a maximum of 49.2% and 50.9%, respectively. The inclusion of EDTA (10 mM), an inhibitor of metalloproteinases, reduced GBM degradation by APMA activated and disrupted PMN by only 7.5%. Incubation of PMN with diphenylene iodonium (DPI) abolished PMN reactive oxygen species generation by > 95% but preserved elastase release. This compound did not directly affect GBM degradation. It did, however, abolish the inhibitory effect of ROS on alpha 1Pi activity and thus indirectly reduced GBM damage by up to 20%.

Basement Membrane↗
PubMed1994

Oxidation of low density lipoprotein by mesangial cells may promote glomerular injury.

Low density lipoprotein (LDL) deposition and local oxidation play a key role in the pathogenesis of atherosclerosis and may likewise contribute to glomerular injury. These studies were designed to determine whether cultured human mesangial cells oxidize homologous LDL and to compare the effects of unmodified and oxidized lipoprotein on cell proliferation, viability and eicosanoid production. Cell-mediated lipoprotein oxidation was demonstrated and could be suppressed by oxygen free radical scavengers and inhibitors of arachidonic acid metabolism. When incubated with cells, oxidized LDL (Ox-LDL) at concentrations up to and including 100 micrograms/ml reduced 3H-thymidine incorporation without causing cytotoxicity as assessed by lactate dehydrogenase release. Under the same conditions there was a concentration-dependent increase in the synthesis of prostaglandins E2,6-keto-PGF1 alpha and thromboxane B2. In contrast, unmodified LDL enhanced DNA synthesis at concentrations less than 40 micrograms/ml and had little effect on eicosanoid production. These results demonstrate that exogenous oxidized LDL inhibits mesangial cell proliferation and increases eicosanoid synthesis. Unmodified lipoprotein can be directly oxidized by these cells through mechanisms that involve generation of oxygen free radicals.

Cell Division↗
PubMed1994

The source and possible significance of hyaluronan in the peritoneal cavity.

The levels of hyaluronan (HA) were determined in dialysate from patients receiving CAPD. The levels found after the four hour dwell were 0.162 +/- 0.08 micrograms/ml (N = 16) in non-infected fluid and 1.69 +/- 1.12 (N = 5) during peritonitis, P < 0.0025. Similarly concentrations in overnight (8 to 10 hours) dwell dialysate were 0.384 +/- 0.22 micrograms/ml (N = 13) when uninfected and 3.17 +/- 2.28 (N = 8) during peritonitis, P < 0.0002. Following de novo catheter insertion the initial peritoneal washout yielded HA levels of 0.0032 +/- 0.0018 micrograms/ml (N = 7). In vitro human peritoneal cells synthesized HA, 90% of which was secreted into the culture medium (CM). By dissociative gel permeation chromatography on Sephacryl 1000 there were no significant differences in the molecular size of HA extracted from (a) non-infected fluid, (b) infected fluid and (c), mesothelial cell culture medium. These results indicate that the process of CAPD results in an increase in HA levels in the peritoneal cavity. During episodes of peritonitis these functions are further augmented. The likely source of HA in the dialysate is the peritoneal mesothelial cells. This response may be an initial event in wound repair.

Adolescent↗
PubMed1994

Identification and independent regulation of human mesangial cell metalloproteinases.

Mesangial cells are known to secrete metalloproteinases that are capable of degrading the constituents of the GBM, and hence are potentially involved not only in the regular maintenance of the ECM components in the glomerulus, but also of contributing to any damage to these components that occurs in disease states. In this report we positively identify by Northern blotting the neutral proteinase that is constitutively secreted by the human mesangial cell (HMC) as gelatinase A (MMP2). Stimulation of HMC gelatinase by IL-1 beta or PMA causes an increase in the total amount of gelatinolytic activity secreted. On examination, however, this increased activity is shown, both by immunoreactivity and by PCR to be due to the induction of the higher molecular weight form of gelatinase, gelatinase B (MMP9), while the amount of gelatinase A remained unaffected. In addition antigen and messenger RNA have been identified for both the specific inhibitors of metalloproteinases TIMP-1 and TIMP-2. The appearance of the larger inducible gelatinase with similar substrate specificity implies that the regular turnover of matrix components may be due to the constitutively released gelatinase A while in pathological situations the inducible gelatinase B becomes predominant. The synthesis and secretion of TIMP-1 and TIMP-2 indicates that the mesangial cell is capable of controlling the activity of its own secreted enzymes.

Base Sequence↗
PubMed1994

Human peritoneal mesothelial cell prostaglandin synthesis: induction of cyclooxygenase mRNA by peritoneal macrophage-derived cytokines.

Increasing evidence suggests that the mesothelial cell contributes to the control of inflammation in both the normal and inflamed peritoneal cavity. The present study examines the regulation of prostaglandin production by human peritoneal mesothelial cells (HPMC) following stimulation with peritoneal macrophage-conditioned medium and the cytokines interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha). IL-1 beta and TNF-alpha stimulated significant release of prostaglandin above background levels in a time and dose dependent manner. Stimulation of HPMC with IL-1 beta (500 pg/ml) or TNF-alpha (100 pg/ml) for 24 hours resulted in the release of 24.5 +/- 4.3 (N = 11) (z = 3.40, P < 0.001 vs. control) and 19.4 +/- 4.5 (N = 10; z = 3.29, P < 0.001 vs. control) pg 6-keto-PGF1 a/micrograms cellular protein, respectively. Pretreatment of HPMC with dexamethasone (10(-6) to 10(-9) M) inhibited both constitutive and cytokine stimulated prostaglandin synthesis in a dose dependent manner. Both PMø-CM and PMø-S.epiCM stimulated 6-keto-PGF1 alpha and PGE2 synthesis by HPMC in a time and dose dependent manner (PMø-S.epiCM >> PMø-CM). Co-incubation of HPMC with PMø-S.epiCM in the presence of anti-IL-1 beta and/or anti-TNF-alpha antibody, interleukin-1 receptor antagonist or soluble TNF receptor (TNF p75) significantly reduced the capacity of these supernatants to stimulate prostaglandin synthesis. Exposure of HPMC to cytokines or PMø-S.epiCM resulted in the time dependent increase in the levels of both Cox-1 and Cox-2 mRNA as assessed by RT/PCR analysis with the greatest increase being seen for Cox-2. These data demonstrate specific stimulation of eicosanoid metabolism in HPMC by peritoneal macrophage derived cytokines, indicating the possible importance of these mediators in the activation of intraperitoneal prostaglandin synthesis. HPMC prostaglandins might act as important pro/anti-inflammatory mediators contributing to a cytokine network in the peritoneal cavity during CAPD peritonitis.

Base Sequence↗