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Biomedical subjects

J F Moorhead

Publications and source records attributed to J F Moorhead.

At least 19 recordsLinked to original sources
PubMed1999

Human mesangial cells express inducible macrophage scavenger receptor.

BACKGROUND: Type A scavenger receptors (Scr) mediate the uptake of modified low-density lipoproteins by macrophages. The accumulation of lipids via this process is thought to lead to foam cell formation in atherosclerotic plaques. Human mesangial cells (HMCs) have not been previously shown to express Scr in normal culture. We therefore investigated whether there is an inducible form of Scr in a human mesangial cell line (HMCL). METHODS: Scr activity was analyzed by cellular uptake of fluorescently labeled acetylated low-density lipoprotein using a flow cytometer. Scr mRNA expression was examined using reverse transcription-polymerase chain reaction, followed by Southern blotting. To investigate the molecular mechanism of Scr expression, several reporter gene constructs were designed. The first contained a full Scr promoter, the second a part of the Scr promoter that has both AP-1 and ets transcription factor binding sites. Other constructs were identical to the second, except that they contained either AP-1 or ets motif mutations. RESULTS: Phorbol 12-Myristate 13-acetate (PMA) and angiotensin II (Ang II) increased both the percentage of Scr-positive cells and the Scr mean fluorescence intensity. PMA and Ang II also increased Scr mRNA and promoter activity in a time- and dose-responsive manner. Protein kinase C and calmodulin transduction pathways were involved in Scr up-regulation induced by PMA and Ang II. Additionally, a serine/threonine kinase was involved in PMA stimulation. Functional analysis showed that both AP-1 and ets motifs were specific response elements to PMA stimulation in HMCLs. CONCLUSIONS: This study suggests that HMCs may express an inducible Scr, by which cells can acquire lipids and convert to foam cells in developing glomerulosclerosis.

Angiotensin II↗
PubMed1999

Lipopheresis in the nephrotic syndrome.

BACKGROUND: Experimental models have established a role for lipoproteins in the pathogenesis of progressive renal failure. However, conventional treatment rarely normalizes the high serum cholesterol of the nephrotic syndrome. The removal of low-density lipoprotein by lipopheresis is discussed. METHODS: Lipopheresis may be beneficial in nephrotic patients with focal segmental glomerulosclerosis. The authors studied the long-term effects of low-density lipoprotein cholesterol (LDL-C) removal using the Kaneka Liposorber system, which binds LDL-C to dextran sulfate in a controlled trial in 20 nephrotic patients with different renal diseases. RESULTS: A 21-month clamp of plasma total cholesterol at 6.0 mmol/liter or below was significantly lower than controls (chi 2 = 84.3, P < 0.001), followed 12 aphereses over 6 to 12 weeks in all but three apheresed patients. 1/Cr slopes were unchanged when the 50-day average period of lipopheresis treatments was excluded from analysis. Proteinuria was not reduced, but serum albumin tended to rise (NS). Fibrinogen fell by 29.8%; high-density lipoprotein, apoA1, and Lp(a) were unchanged. Two apheresed patients had a prolonged remission with a reduction of proteinuria to less than 250 mg/24 hr. The reasons for prolonged reduction of total cholesterol include depletion of tissue cholesterol, an improved fractional catabolic rate of very low density lipoprotein (VLDL), increased hepatocyte LDL turnover, and the maintenance of statin therapy. CONCLUSION: Lipopheresis is a safe and effective method for the control of LDL in nephrotic syndrome. Early clamping of total cholesterol in the normal range resulted in a prolonged and significant reduction of LDL compared with controls.

Anticholesteremic Agents↗
PubMed1999

Calcineurin inhibitors enhance low-density lipoprotein oxidation in transplant patients.

BACKGROUND: Our objective was to assess the pro-oxidant status of neoral and tacrolimus in renal transplant patients and monitor the protection provided by vitamin C and vitamin E in normalizing low density lipoprotein (LDL) oxidation lag time of tacrolimus-treated patients. METHODS: Plasma LDL was isolated by density gradient ultracentrifugation from renal transplant patients receiving neoral, tacrolimus and tacrolimus with vitamin C and vitamin E. Oxidation was initiated by the addition of CuCl2 at 37 degrees C and monitored at 234 nm over 480 minutes and oxidation lag time was computed. Total antioxidant capacity of serum was measured using the enhanced chemiluminescent method. RESULTS: LDL from tacrolimus-treated patients had significantly lower oxidation lag time and serum antioxidant activity in comparison with neoral-treated patients, and this was particularly significant during the first four months after transplantation. Vitamin C and E supplementation in tacrolimus treated patients provided protection against oxidation and normalized their oxidation lag time. CONCLUSION: Calcineurin-inhibiting drugs, CsA and tacrolimus, have pro-oxidant activity and they increase the susceptibility of LDL to oxidation. Neoral formulation is fortified with DL-alpha tocopherol and therefore provides protection against oxidation. The present study clearly demonstrates the benefit of giving vitamin C and E supplements to patients taking tacrolimus and this seems to be particularly important during the early period after transplantation.

Adult↗
PubMed1999

Lysophosphatidylcholine induces platelet-derived growth factor gene expression in a human mesangial cell line.

BACKGROUND: Oxidized low-density lipoprotein (oxLDL) has been considered important in the pathogenesis of progressive renal injury. Lysophosphatidylcholine (lysoPC) is a major phospholipid component of oxLDL. On the other hand, platelet-derived growth factor (PDGF) has also been implicated in proliferative disease of the kidney. This study investigated the difference in the potential of PC and lysoPC to induce DNA synthesis and PDGF gene expression in a human glomerular mesangial cell line (HMCL). METHODS: DNA synthesis in HMCL was measured by [3H] thymidine incorporation. The mRNA expression levels of the PDGF A chain and B chain genes were measured using reverse transcription-polymerase chain reaction. RESULTS: LysoPC treatment up-regulated the [3H] thymidine incorporation level in a dose-dependent fashion. The [3H] thymidine incorporation level in HMCL coincubated with lysoPC started to increase after 4 hours of treatment, peaked at 24 hours, and decreased thereafter. The level in HMCL incubated with 100 microM of lysoPC (palmitoyl or stearoyl) increased to 7- or 10-fold of the control at peak time, respectively. However, PC treatment did not increase [3H] thymidine incorporation in HMCL. PC treatment did not induce mRNA expression of either PDGF A or B chain genes. LysoPC did not induce PDGF A chain mRNA expression either. The only B chain mRNA expression was induced by lysoPC. The mRNA expression level in HMCL treated with 50 microM lysoPC for two hours increased to 1.6-fold that of the control. CONCLUSION: LysoPC may induce DNA synthesis in a mesangial cell through the induction of PDGF BB as an autocrine and paracrine growth factor.

Cell Line↗
PubMed1999

Renal vasoconstriction induced by oxidized LDL is inhibited by scavengers of reactive oxygen species and L-arginine.

BACKGROUND: Low density lipoprotein (LDL) may be involved in the pathogenesis of glomerulosclerosis and progressive renal dysfunction associated with atherosclerotic renal artery stenosis (RAS). This study was undertaken to investigate the effects of native (n-LDL) and oxidized LDL (ox-LDL) on renal vascular response and function in an isolated perfused rat kidney (IPRK) model. MATERIAL AND METHOD: IPRK model was used for the study at a constant pressure of 100 mm of Hg in the renal artery with continuous monitoring of pressure and renal perfusate flow. Urine and perfusate samples were collected to determine [14C] Inulin clearance and fractional reabsorption of sodium. To elucidate the role of nitric oxide (NO) urinary c-GMP, nitrate and nitrite excretion were measured and the responses to the NO synthase inhibitor N-monomethyl-L-arginine (LNMMA) and the NO donor Nitroso-glutathione (GSNO) were assessed. The effect of L-arginine supplementation and the role of reactive oxygen species were also studied by adding superoxide dismutase (SOD) and catalase. RESULTS: Ox-LDL but not n-LDL caused vasoconstriction in IPRK, as evidenced by a significant dose dependent reduction in renal perfusate flow. [14C] Inulin clearance and fractional reabsorption of sodium were reduced during ox-LDL infusion whereas no significant change occured with n-LDL. There was a significant decrease in urinary excretion of c-GMP during ox-LDL infusion. 10 microM LNMMA significantly increased and GSNO (10 microM) significantly diminished the vasoconstrictory effect of ox-LDL. The presence of L-arginine (100 & 500 microM) significantly decreased ox-LDL induced vasoconstriction. SOD (150 U/ml) and catalase (1200 U/ml) both had a significant inhibitory effect and the combination of SOD and catalase almost completely abolished the vasoconstriction due to ox-LDL. CONCLUSION: These results suggest that ox-LDL induced vasoconstriction in IPRK is mediated by decreased activity of NO probably due to inactivation of NO by reactive oxygen species. The free radical scavengers SOD, catalase and L-arginine provided protection against ox-LDL induced vasoconstriction in this model.

Animals↗
PubMed1999

LDL receptor gene expression in human mesangial cells under the influence of calcium channel blockers.

BACKGROUND: Intracellular transport of lipid through the regulation of LDL receptor (LDLr) may be important in the progression of renal dysfunction. METHODS: We explored LDLr gene expression in human mesangial cell line (HMCL) under influence of calcium channel blockers using cell proliferation, LDL binding, Northern blot and LDLr promoter activity assay. RESULTS: Diltiazem and verapamil increased the expression of LDLr mRNA in a dose-dependent manner. Increased LDLr mRNA paralleled LDL binding. Nifedipine did not increase the expression of LDLr mRNA and LDL binding to HMCL at 1 - 100 micromol/l. The LDLr promoter activity assay showed that treatment with 100 micromol/ of diltiazem and verapamil increased LDLr promoter activity by 126.72 +/- 10.68%, and 166.41 +/- 11.41%, respectively, at 24 hours (control as 100%), while treatment with 100 micromol/l of nifedipine had an inhibitory effect on LDLr promoter activity. High concentration of LDL (250 microg/ml) inhibited promoter activity. Diltiazem or verapamil coincubated with LDL (250 microg/ml) could not override transcriptional inhibition by LDL. CCBs inhibited the proliferation of HMCL, therefore, CCBs-induced LDLr expression did not depend on a proliferative response. Signal transduction pathway experiments showed that the calmodulin transduction pathway was involved in LDLr upregulation induced by diltiazem or verapamil. Additionally, tyrosine kinase and PKC pathways were involved in the induction of LDLr induced by verapamil. CONCLUSION: These studies show that diltiazem and verapamil increase LDLr gene transcription and expression which is independent of cell proliferation in HMCL.

Blotting, Northern↗
PubMed1999

Human mesangial cells express inducible macrophage scavenger receptor: an Ap-1 and ets mediated response.

BACKGROUND: Type A scavenger (SR) mediate the uptake of modified low-density lipoproteins by macrophages. The accumulation of lipid via this process is thought to lead to foam cell formation in atherosclerotic plaques. Human mesangial cells (HMC), which can be converted to foam cells in vivo, have not previously been shown to express SR in normal culture. We investigated whether or not there was an inducible form of SR in a human mesangial cell line (HMCL). METHODS: SR activity was analyzed by cellular uptake of fluorescently labeled acetylated low-density lipoprotein using a flow cytometer. SR mRNA expression was examined using RT-PCR followed by Southern blotting. To investigate the molecular mechanism of SR expression, several reporter gene constructs were designed. The first contained a full SR promoter, the second a part of the SR promoter that has both activated protein-1 (AP-1) and ets transcriptional factor binding sites. Other constructs were identical to the second except they contained either AP-1 or ets motif mutations. RESULTS: Phorbol 12-myristate 13-acetate (PMA) increased both the percentage of SR positive cells and SR mean fluorescence intensity. PMA also increased SR mRNA and promoter activity in a time and dose responsive manner. Function analysis showed that both AP-1 and ets motifs were specific response elements to PMA stimulation in HMCL. CONCLUSIONS: The present study suggests that the combination of interaction between AP-1 and ets transcriptional factors may mediate the inducible expression of the SR gene in HMCL, which may contribute to foam cell formation.

Binding Sites↗
PubMed1998

Differential ability of cells to promote oxidation of low density lipoproteins in vitro.

Atherosclerosis and focal segmental glomerulosclerosis share some common histological features and it is speculated that they result from similar pathobiological mechanisms. There is strong evidence that oxidation of low density lipoprotein (LDL) may be an initiating event in atherogenesis and that oxidised LDL may also be involved in the glomerulosclerotic process. In vitro studies have demonstrated that cells present in the arterial intima and kidney-derived cells promote LDL oxidation. The aim of this study was to compare LDL oxidation by kidney-derived human mesangial cells and proximal tubular cells, with human umbilical vein endothelial cells and the human monocyte cell line THP-1. We used the thiobarbituric acid assay and agarose gel electrophoresis to measure the extent of LDL oxidation. Our results demonstrate that all of the cell types used had the ability to oxidise LDL significantly more than cell-free controls and that endothelial cells induced the highest degree of oxidative modification of LDL under our experimental conditions.

Arteriosclerosis↗
PubMed1998

Cytokine regulation of low-density lipoprotein receptor gene transcription in human mesangial cells.

BACKGROUND: The intracellular transport of lipids through regulation of the LDL receptor (LDLr) may be important in the progression of renal dysfunction. The present study was undertaken to investigate whether cytokines have any major effects on LDLr regulation and lipid-mediated glomerular injury in human mesangial cells (HMC). METHODS: We explored the effects of 50 ng/ml of tumour necrosis factor alpha (TNF alpha), 5 ng/ml of transforming growth factor beta (TGF beta), platelet-derived growth factor (PDGF), and interleukin-1beta (IL-1beta) on the regulation of LDLr gene transcription in a human mesangial cell line (HMCL) using cell proliferation, LDL binding, northern blot and LDLr promoter activity assays. RESULTS: TNF alpha, TGF beta, PDGF or IL-1beta did not significantly stimulate HMCL proliferation at the concentrations given above, but maximally stimulated LDLr mRNA expression and increased LDLr promoter activity by 167.48+/-23.56%, 150.47+/-24.41%, 127.71+/-24.65% and 163.01+/-31.91% respectively, at 24 h. An increased LDL binding was observed in parallel with increased LDLr mRNA. The tyrosine kinase transduction pathway was involved in LDLr upregulation induced by all four cytokines. Additionally, TGF beta involved serine/threonine kinase and G-protein pathways, and IL-1beta involved calmodulin, serine/threonine kinase and PKC pathways in upregulating LDLr. A high concentration of LDL (250 microg/ml) inhibited promoter activity, but TNF alpha, TGF beta, PDGF and IL-1beta co-incubated with LDL could override transcriptional inhibition by LDL. CONCLUSION: TNF alpha, TGF beta, PDGF and IL-1beta increased LDLr gene expression by increasing sterol-independent and mitogenesis-independent gene transcription. This process may contribute to lipid deposition and foam cell formation in HMC.

Base Sequence↗
PubMed1997

Glomerular atherosclerosis.

This paper suggests a two-hit model for lipoprotein-mediated progressive renal disease, in which postsecretory modification of low-density lipoprotein may favour the transformation of mesangial cells, monocytes and macrophages to glomerular foam cells. Proteinuria and lipiduria would mediate tubulointerstitial damage. Based on this, careful treatment with lipid-lowering agents, lipopheresis and antioxidants may ameliorate the progression of glomerular and tubulointerstitial pathology.

Arteriosclerosis↗
PubMed1996

Do glomerular atherosclerosis and lipid-mediated tubulo-interstitial disease cause progressive renal failure in man?

The nephrotic syndrome presents the kidney with a new environment in which blood vessels, glomerular structures and tubules are exposed over substantial periods of time to lipoproteins. LDL has charge affinity with glomerular basement membrane glycosaminoglycans, so potentially increases or maintains albumin loss. This in turn stimulates LDL synthesis. HDL is small enough to be passed by the glomerular filter in substantial amounts and has been found to stimulate endothelin-1 production by human proximal tubular cells in culture. LDL also inhibits nitric oxide vasodilatory responses, an action which when added to that of endothelin-1 may result in decreased renal tissue oxygenation. Taken together, these aspects of the nephrotic syndrome broaden conventional definitions of atherosclerosis and offer a number of targets for therapy in progressive renal disease.

Arteriosclerosis↗
PubMed1995

Cost-effective rehabilitation: Part 1--Basic principles.

Patients may be referred for rehabilitation services with a variety of diagnoses, including stroke, spinal cord injury, brain injury, multiple trauma, orthopedic disorders, arthritis, multiple medical problems, and chronic pain. The goals and endpoints for treatment of these conditions are often unclear. The principles that are described in this article provide a structure for creating efficient and effective rehabilitation treatment plans. These principles can also focus the utilization and peer review processes and can assist in determining medical necessity of rehabilitation services. Part one of this two-part article discusses general principles of cost-effective rehabilitation. Part two, in the December 1995 issue, will focus on individual patient management issues.

Cost-Benefit Analysis↗
PubMed1995

Cost-effective rehabilitation: Part 2--Approaches to patient management problems.

Part one of this two-part series discussed general principles of cost-effective rehabilitation: Patients in rehabilitation programs should be working toward achievement of real-world functional goals. Goals should be realistic, and reachable in a reasonable amount of time. Rehabilitation services should be provided at the lowest safe and effective level of care appropriate to the patient's needs. Patients should be participating to their full potential in an active therapy program. Therapy intended to maintain a patient's current condition should be carried out by nonprofessionals who have had training sessions with rehabilitation professionals as needed. Discharge planning begins on the day of admission to the inpatient or outpatient rehabilitation program. This second of the two-part series will focus on individual patient management issues. It discusses circumstances in which the principles of cost-effective rehabilitation may need to be modified. It also discusses approaches to remedy patient management problems that may lead to excessive or ineffective utilization of rehabilitation services.

Case Management↗
PubMed1994

Tubular lipidosis: epiphenomenon or pathogenetic lesion in human renal disease?

Tubular lipidosis is a commonly observed histological lesion in proteinuric renal diseases. We have studied the interaction between native and modified human lipoproteins and human renal proximal tubular cells to investigate whether lipoproteins could be injurious to tubular cells in culture. Human renal proximal tubular cells were cultured and characterized by established methods. Preliminary studies showed that these cells could take up and degrade normal human lipoproteins by high affinity (HDL) and low affinity (LDL) pathways. In subconfluent culture, native lipoproteins, that is, LDL, HDL2 and HDL3, had markedly different effects on cell growth as measured by 3H-thymidine uptake and total cell protein as compared to modified lipoproteins such as minimally modified and oxidized LDL. In addition, we found that renal tubular cells could oxidized native LDL in the presence of copper largely by a superoxide-mediated mechanism. Finally, cellular accumulation of lipid was demonstrated in vitro by incubating cultured cells with varying lipoprotein concentrations for up to 48 hours. Notably, cell detachment was observed only with high concentrations of modified LDL especially with minimally modified LDL. We speculate that uptake and oxidation of filtered LDL by tubular cells may lead to tubular injury in nephrotic states.

Cell Division↗
PubMed1994

Human high density lipoproteins stimulate endothelin-1 release by cultured human renal proximal tubular cells.

The vasoconstrictive and mitogenic actions of endothelins have been implicated in the pathogenesis of progressive renal disease. In the present study, we have assessed whether plasma high density lipoproteins (HDL), the major filtered urinary lipoprotein in nephrotic states, can influence endothelin-1 (ET-1) production by cultured human renal proximal tubular cells. Human HDL was found to stimulate ET-1 secretion up to fourfold in a dose- and time-dependent manner; the effect was greater in subconfluent cultures than in confluent ones. There was little difference between the stimulatory effect of HDL2 and the major HDL subclass, HDL3. Preincubation of the cells with albumin did not abolish the HDL effect, while partially- or fully-delipidated HDL3 largely reproduced the effect of whole HDL3. These findings suggest that stimulation of ET-1 secretion was not simply due to protein or lipid repletion of the cells. Rather, the effect was mediated by HDL apolipoproteins, although binding to the HDL receptors involved in cellular cholesterol homeostasis was not required as tetranitromethane-modified HDL3 was an equally effective agonist of ET-1 release. Apolipoprotein (apo) A-I was indirectly implicated in the process since modified HDL3 in which apoA-II largely replaced apoA-I was less potent than HDL3. A one hour exposure of the cells to HDL3 was sufficient to activate ET-1 production for the following 12 hours, although maximum activation required six hours.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaloids↗