No abstract available.

No abstract available.
Brain Stem Infarctions*

BACKGROUND: High glucose upregulates MCP-1 expression in rat glomerular mesangial cells and in human peritoneal mesothelial cells. However, the role of high glucose-induced MCP-1 on the development and progression of diabetic renal injury and peritoneal injury during peritoneal dialysis(PD) using high glucose PD solutions are not clear. Since MCP-1 was shown to upregulate transforming growth factor-beta1(TGF-beta1) and collagen expression in lung fibroblasts, the present study investigated the effects of MCP-1 on fibronectin secretion by mouse mesangial cells(MMC), human peritoneal mesothelial cells (HPMC), and human peritoneal fibroblasts(HPFB). METHODS: Synchronized cells were stimulated by different concentrations of MCP-1(0.1-100 ng/mL) or TGF-beta1(0.1-10 ng/mL) for 48 hours. Fibronectin protein secreted into the media was analyzed by Western blot analysis. RESULTS: MCP-1 up to 100 ng/mL did not affect fibronectin secretion by MMC. TGF-beta1 10 ng/mL, however, increased fibronectin secretion by MMC 2.8 fold that of control. MCP-1 up to 100 ng/mL did not affect fibronectin secretion by HPMC. But, TGF-beta1 0.1 ng/mL increased fibronectin secretion by HPMC 1.8 fold compared to control. On the other hand, MCP-1 increased fibronectin secretion by HPFB in a dose-dependent manner. MCP-1 at 1-10 ng/mL significantly increased fibronectin when compared to M199 control. 100 ng/mL MCP-1 further increased fibronectin secretion by HPFB compared to 0.1-10 ng/mL MCP-1. CONCLUSION: These results suggest a possible role for MCP-1 in the development and progression of peritoneal fibrosis and support the view that in addition to recruiting inflammatory cells MCP-1 may play a role in tissue fibrosis in other organs.
Animals ; Blotting, Western ; Chemokine CCL2* ; Collagen ; Fibroblasts* ; Fibronectins* ; Fibrosis ; Glucose ; Hand ; Humans* ; Lung ; Mesangial Cells ; Mice ; Monocytes* ; Peritoneal Fibrosis ; Rats ; Transforming Growth Factor beta1

BACKGROUND: Advanced glycation end products (AGE) are independent risk factors in the development and progression of diabetic nephropathy. Receptor for AGE(RAGE) is considered the main receptor involved in AGE-induced cell activation. Galectin-3, another AGE receptor, has recently been found upregulated in mesangial cells(MC) cultured under high glucose and in diabetic rat kidneys. N epsilon(carboxymethyl)lysine(CML) is a well characterized AGE but its role in MC activation is unknown. The present study examined the effects of CML on MC proliferation and extracellular matrix(ECM) secretion. METHODS: Synchronized rat MC were stimulated with different concentrations of CML-bovine serum albumin(BSA), control BSA, and transforming growth factor-beta(TGF-beta) for up to 72 hours. Cell proliferation was measured by [3H]-thymidine incorporation. Fibronectin, TGF-beta, plasminogen activator inhibitor(PAI)-1 secreted into the media and RAGE and galectin-3 expression in MC were measured by Western blot analysis and ELISA. RESULTS: 1,000 micro /mL of CML-BSA decreased [3H]-thymidine incorporation by MC at 48 hours and 10 ng/mL TGF-beta at 24 and 48 hours. CML-BSA 100 and 1,000 micro /mL, control BSA 1,000 micro /mL, and TGF-beta 10 ng/mL increased fibronectin secretion at 48 hours. CML-BSA up to 1,000 micro /mL did not affect TGF-beta or PAI-1 secretion. TGF-beta 10 ng/mL, however, significantly increased PAI-1 secretion. Cultured MC expressed both RAGE and galectin-3. CML-BSA 100 micro /mL upregulated galectin-3 expression. CONCLUSION: CML-BSA decreased MC proliferation and increased fibronectin secretion, suggesting that CML may lead to ECM accumulation and glomerulosclerosis in diabetic animals. MC express RAGE and galectin-3 constitutively and CML-induced galectin-3 upregulation may have a role in AGE-induced MC activation.
Animals ; Blotting, Western ; Cell Proliferation ; Diabetic Nephropathies ; Enzyme-Linked Immunosorbent Assay ; Fibronectins ; Galectin 3 ; Glucose ; Glycosylation End Products, Advanced ; Kidney ; Mesangial Cells* ; Plasminogen Activator Inhibitor 1 ; Plasminogen Activators ; Rage ; Rats ; Risk Factors ; Transforming Growth Factor beta ; Up-Regulation ; Advanced Glycosylation End Product-Specific Receptor

Preservation of peritoneal membrane function is important in the success of long-term peritoneal dialysis (PD). During PD, human peritoneal mesothelial cells (HPMC) are continuously exposed to unphysiological peritoneal dialysis solution(PDS) charaterized by high glucose and lactate concentrations, low pH, and hyperosmolality. Since few studies have examined the effects of lactate and pH on HPMC biology, the present study investigated the effects of lactate and pH on the viability and proliferation of cultured HPMC and on the production of TGF-beta1, a fibrogenic cytokine, and fibronectin by cultured HPMC. HPMC were obtained from the omental tissue of pregnant women who were undergoing Cesarean section. Cells at confluence were utilized to determine the viability(LDH release), proliferation([3H]-thymidine incorporation), and the production of fibronectin and TGF-beta1(ELISA) after synchronizing the cell growth by incubating with serum free media for 24 hours. After exposure to the media containing lactate and pH, LDH release increased in dose- and time-dependent manner. Both 1.5% and 4.25% commercial PD solutions were cytotoxic and induced more than 80% LDH release within 24 hours. LDH release decreased with increasing dilution of commercial peritoneal dialysate, but there was no significant difference in LDH release between 1.5% and 4.25% PDS. LDH release increased in response to pH 5.5. Thymidine incorporation assay revealed that lactate and low pH significantly inhibited proliferation of HPMC. ELISA revealed that exposure of HPMC to lactate and low pH decreased fibronectin protein synthesis, when compared to cell exposed to bicarbonate containing M199 media. Our results clearly show that lactate and low pH lead to dose- and time-dependent cell death and reduce proliferation of cultured HPMC. Lactate and low pH per se appear to decrease fibronectin production by HPMC but may set a stage for other factors to promote progressive fibrosis during the healing stage in long-term PD.
Biology ; Cell Death ; Cesarean Section ; Culture Media, Serum-Free ; Enzyme-Linked Immunosorbent Assay ; Female ; Fibronectins ; Fibrosis ; Glucose ; Humans* ; Hydrogen-Ion Concentration* ; Lactic Acid* ; Membranes ; Peritoneal Dialysis ; Pregnancy ; Pregnant Women ; Thymidine ; Transforming Growth Factor beta1

BACKGROUND: 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a prototype compound of polyhalogenated aromatic hydrocarbons, produces diverse biologic effects. Although nephrotoxicity of aromatic hydrocarbons such as benzo[a]pyrene(BP) is well known, little is known about the effects of TCDD on renal function. Thus, the present study examined the effects of TCDD on cell viability, proliferation, and extracellular matrix(ECM) synthesis by glomerular mesangial cells, LLC-PK1 cells representing proximal tubular epithelial cells, and MDCK cells representing distal epithelial cells and compared with the effects of BP. METHODS: Quiescent cells were incubated with serum free media containing different concentrations of TCDD(1-100 nM) and BP(3 and 30 micro M) for 24- 96 hours. Cell viability and proliferation were assessed by lactate dehydrogenase(LDH) release and [3H]-thymidine incorporation, respectively. Secreted fibronectin was measured by Western blot analysis. RESULTS: When cells were continuously exposed to TCDD, LDH release significantly increased in MMC, LLC-PK1, and MDCK in a dose- and a time- dependent manner. [3H]-Thymidine incorporation was increased in MMC and LLC-PK1 but decreased in MDCK by TCDD. Contrary to TCDD, 30 micro BP significantly inhibited [3H]-thymidine incorporation in MMC and MDCK but not in LLC-PK1. Both TCDD and BP increased fibronectin secretion by MMC, LLC-PK1, and MDCK cells, suggesting that TCDD and BP may cause renal fibrosis leading to loss of renal function. CONCLUSION: These data provide experimental evidence that TCDD can alter cell viability and proliferation and increase ECM synthesis by renal cells which may lead to renal injury.
Animals ; Blotting, Western ; Cell Survival ; Culture Media, Serum-Free ; Epithelial Cells* ; Fibronectins ; Fibrosis ; Hydrocarbons, Aromatic ; Lactic Acid ; LLC-PK1 Cells ; Madin Darby Canine Kidney Cells ; Mesangial Cells ; Swine ; Tetrachlorodibenzodioxin

Human chorionic gonadotropin (HCG) is one of the glycoproteins families synthesized by the placenta, and consists of 2 noncovalently joined subunits, namely, alpha and beta. The alpha and beta-subunits have a structural homology with the alpha and beta-subunits of TSH and LH. The thyrotropic action of HCG results from its structural similarity to TSH, so beta-HCG can bind to the TSH receptor in the thyroid gland. A high level of HCG, accompanied by an increased thyroid hormone level, can be observed in gestational trophoblastic diseases (GTD), such as a hydatidiform mole or a choriocarcinoma. However, the clinical symptoms of hyperthyroidism in GTD are rarely observed. A 27-years-old woman, admitted due to an amenorrhea of 11 weeks duration, with thyrotoxic symptoms, such as weight loss, palpitation, sweating, tremor, heat intolerance and anxiety, was evaluated. Her serum free T4 level was 8 times higher than normal, and her serum beta-HCG level was over 1,000,000IU/L. She had a curettage operation, with the pathological findings of a complete hydatidiform mole. These thyrotoxic symptoms developed due to a hydatidiform mole, and were accompanied with a highly increased serum beta-HCG level. After evacuation of the molar tissue, the thyroid hormone and thyrotoxic symptoms normalized. Here, this case is reported, with brief review of the literature.
Amenorrhea ; Anxiety ; Choriocarcinoma ; Chorionic Gonadotropin ; Curettage ; Female ; Gestational Trophoblastic Disease ; Glycoproteins ; Hot Temperature ; Humans ; Hydatidiform Mole* ; Hyperthyroidism ; Molar ; Placenta ; Pregnancy ; Receptors, Thyrotropin ; Sweat ; Sweating ; Thyroid Gland ; Thyrotoxicosis* ; Tremor ; Weight Loss

BACKGROUND/AIMS: The relationship between Runt-related transcription factor 3 (RUNX3) gene inactivation and various solid tumors has been reported; however, little information is available about RUNX3 in thyroid cancers. METHODS: We evaluated the DNA methylation of RUNX3 in 13 papillary thyroid cancer tissues and four thyroid cancer cell lines. Additionally, using reverse transcriptase-polymerase chain reaction, we analyzed RUNX3 gene expression in several thyroid cancer cell lines after treating with the demethylating agent 5-aza-2'-deoxycytidine (DAC). RESULTS: RUNX3 was hypermethylated in many thyroid cancer cell lines and in 10 of the 12 papillary thyroid cancer tissues. Treatment with DAC increased the expression of RUNX3 in some thyroid cancer cell lines. CONCLUSIONS: We suggest that RUNX3 is associated with thyroid carcinogenesis, and RUNX3 methylation is a potentially useful diagnostic marker for papillary thyroid cancer.
Azacitidine/analogs & derivatives/pharmacology ; Carcinoma/*genetics ; Cell Line, Tumor ; Core Binding Factor Alpha 3 Subunit/*genetics ; DNA Methylation/drug effects ; Gene Expression/drug effects ; Humans ; Thyroid Neoplasms/*genetics ; Tumor Markers, Biological/genetics