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Biomedical subjects

S Yung

Publications and source records attributed to S Yung.

At least 19 recordsLinked to original sources
PubMed2006

Increased mesangial cell hyaluronan expression in lupus nephritis is mediated by anti-DNA antibody-induced IL-1beta.

The mechanism by which anti-DNA antibodies contribute to the pathogenesis of lupus nephritis (LN) remains to be fully elucidated. Hyaluronan (HA) is an important extracellular matrix constituent that accumulates during tissue injury, and participates in lymphocyte recruitment to sites of inflammation. The role of HA in the pathogenesis of LN has not been defined. We investigated the expression of HA in renal biopsies and circulating HA levels in patients with diffuse proliferative LN, and the effect of human anti-DNA antibodies on HA synthesis in cultured human mesangial cells (HMC). HA expression was increased in the mesangium, and in the periglomerular and tubular distribution in LN kidney biopsies. LN patients showed increased levels of circulating HA, especially during active disease, which correlated with anti-DNA antibody titers (r=0.35, P=0.0234). Anti-DNA antibodies isolated during active LN but not remission increased de novo synthesis of (3)H-labeled HA, which was accompanied by induction of HA synthase (HAS) II transcription, and enhanced IL-1beta, IL-6, and tumor necrosis factor-alpha secretion in HMC (P<0.001 for all). Only anti-DNA antibody induction of IL-1beta enhanced HA synthesis, which was abrogated by inhibitors of de novo mRNA or protein synthesis. Our findings demonstrate that HA expression is significantly increased within the mesangium in diffuse proliferative LN mediated through anti-DNA antibody-induced IL-1beta. Given that HA plays a pivotal role during inflammatory responses, influences cellular behavior and assists in the recruitment of lymphocytes to sites of injury, it is likely that HA contributes to the pathogenesis of LN.

Adult↗
PubMed2001

Analysis of apicoplast targeting and transit peptide processing in Toxoplasma gondii by deletional and insertional mutagenesis.

Deletion and insertion mutagenesis was used to analyze the targeting sequence of the nuclear encoded apicoplast protein, the ribosomal protein small subunit 9 of Toxoplasma gondii. Previous studies have shown that nuclear encoded apicoplast proteins possess bipartite leaders having characteristic signal sequences followed by serine/threonine rich transit sequences. Deletion analysis demonstrated that the first 55 amino acids of the rps9 leader were sufficient for apicoplast targeting. Insertional mutagenesis tagging the leader sequence with a hemagglutinin (HA) tag was used to study the events involved in the targeting pathway. Transfectants with insertions near the N-terminus of the transit displayed HA tagged precursors outside of the apicoplast, in the perinuclear region. In contrast, transfectants with the HA tag inserted near the carboxyl end of the transit-like region had apicoplast labeling. Western blot analysis of HA tagged stable isolates suggested that processing of the HA tagged leaders was a multi-step process, with processing occurring both outside of and at or within the apicoplast.

Animals↗
PubMed2001

Emodin ameliorates glucose-induced morphologic abnormalities and synthesis of transforming growth factor beta1 and fibronectin by human peritoneal mesothelial cells.

OBJECTIVE: Excessive synthesis and deposition of matrix proteins by peritoneal mesothelial cells can lead to structural and functional changes in the peritoneal membrane, jeopardizing the long-term efficacy of peritoneal dialysis (PD). Prolonged exposure to high glucose concentrations in PD fluid has been implicated as a major stimulus to matrix accumulation, through the induction of transforming growth factor beta1 (TGFbeta1). This study investigated the effect of emodin (3-methyl-1,6,8-trihydroxyanthraquinone) on TGFbeta1 and fibronectin (FN) synthesis in human peritoneal mesothelial cells (HPMCs) under high glucose concentration. DESIGN: The HPMCs were preconditioned in either 5 mmol/L or 30 mmol/L D-glucose for 2 weeks prior to the addition of emodin. Cell viability was assessed by MTT assay and lactate dehydrogenase (LDH) release. Morphology of HPMCs was studied by phase-contrast microscopy. Modulation of TGFbeta1 and FN synthesis at transcription and translation were investigated by reverse transcriptase polymerase chain reaction (RT-PCR), ELISA, and Western blot analysis. RESULTS: When cultured under 30 mmol/L D-glucose, HPMCs demonstrated increased cell volume, multinucleation, and denudation of the monolayer, as compared with cells cultured under a physiologic (5 mmol/L) glucose concentration. High glucose concentration induced TGFbeta1 synthesis by HPMCs (217.17 +/- 14.88 pg/mL at 5 mmol/L D-glucose vs 370.33 +/- 20.67 pg/mL at 30 mmol/L D-glucose, p < 0.0001), and FN synthesis was induced at transcription and translation. Mannitol at 30 mmol/L did not affect HPMC morphology; matrix synthesis was also unaltered. Administration of emodin together with 30 mmol/L D-glucose resulted in amelioration of cell enlargement and exfoliation, and abrogation of TGFbeta1 induction (370.33 +/- 20.67 pg/mL for 30 mmol/L D-glucose alone vs 260.50 +/- 17.89 pg/mL for 30 mmol/L D-glucose + emodin, p < 0.0001). Synthesis of FN induced by high glucose was also reduced by 40% in the presence of emodin. CONCLUSIONS: These findings provide the first evidence that emodin can ameliorate high glucose-induced matrix synthesis in HPMCs by suppression of TGFbeta1. Emodin may thus be useful in preserving peritoneal integrity in PD.

Blotting, Western↗
PubMed2000

Induction of hyaluronan metabolism after mechanical injury of human peritoneal mesothelial cells in vitro.

BACKGROUND: Hyaluronan (HA) is an important extracellular matrix component that is involved in cell movement and tissue repair. In vertebrates, HA synthase genes (HAS 1, HAS 2, and HAS 3) that control the synthesis of HA have been identified. In this article, we investigated HA synthesis in the response of human peritoneal mesothelial cells (HPMCs) to injury. METHODS: The expression of HAS 1, HAS 2, and HAS 3 mRNA and the synthesis of [(3)H]-labeled HA were examined in an in vitro model of peritoneal mesothelial cell damage. The staining for uridine diphosphoglucose dehydrogenase, a key enzyme in the synthesis of HA, and biotinylated HA-binding protein was used to determine the cellular location of HA synthesis and its site of deposition. RESULTS: Growth-arrested human HPMCs expressed low levels of mRNA for HAS 2 and HAS 3 but not HAS 1. Following injury to the monolayer, HAS 2 was up-regulated by 6 hours, reaching maximal expression between 12 and 24 hours. In contrast, the expression of HAS 3 was down-regulated. During the same time period, synthesis of HA was increased in the injured monolayer. This synthetic activity appeared to be restricted to cells at the edge of the wound and to cells entering the wound. In a separate series of experiments, the addition of HA to the injured monolayer at a concentration range found in peritoneal fluid (50 to 3300 ng/mL) increased the migration of cells into the wound in a dose-dependent manner. CONCLUSIONS: These studies provide evidence that HA is an important component of peritoneal mesothelial cell migration. The results also suggest that in this process, there is differential regulation of HAS gene expression and that the synthesis of HA is limited to cells located at the leading edge of the wound.

Cell Movement↗
PubMed2000

Neuroendocrine mechanisms for reproductive senescence in the female rat: gonadotropin-releasing hormone neurons.

Reproductive aging in female rats is characterized by profound alterations in the neuroendocrine axis. The preovulatory luteinizing hormone (LH) surge is attenuated, and preovulatory expression of the immediate early gene fos in gonadotropin-releasing hormone (GnRH) neurons is substantially reduced in middle-aged compared with young rats. We tested the hypothesis that alterations in GnRH gene expression may be correlated with the attenuation of the LH surge and may be a possible mechanism involved in neuroendocrine senescent changes. Sprague-Dawley rats ages 4 to 5 mo (young), 12-14 mo (middle-aged), or 25 to 26 mo (old) were killed at 10:00 AM or 3:00 PM on proestrus, the day of the LH surge, or diestrus I in cycling rats, and on persistent estrus or persistent diestrus in acyclic rats. RNase protection assays of GnRH mRNA and GnRH primary transcript were performed. GnRH mRNA levels increased significantly with age, whereas GnRH primary transcript levels, an index of GnRH gene transcription, decreased in old compared to young and middle-aged rats. This latter result suggests that an age-related change in GnRH mRNA levels occurs independently of a change in gene transcription, indicating a potential posttranscriptional mechanism. On proestrus, GnRH mRNA levels increased significantly from 10:00 AM to 3:00 PM in young rats. This was in contrast to proestrous middle-aged rats, in which this afternoon increase in GnRH mRNA levels was not observed. Thus, the normal afternoon increase in GnRH mRNA levels on proestrus is disrupted by middle age and may represent a substrate for the attenuation of the preovulatory GnRH/LH surge that occurs in rats of this age, prior to reproductive failure.

Aging↗
PubMed1998

Bikunin present in human peritoneal fluid is in part derived from the interaction of serum with peritoneal mesothelial cells.

We recently reported that peritoneal fluid mainly contains two proteoglycans; one is the interstitial proteoglycan referred to as decorin, and the other an uncharacterized small chondroitin sulfate proteoglycan. In the present study, we have used a two-step process to isolate the small chondroitin sulfate proteoglycan free of decorin. The purified molecule ran as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with apparent molecular mass 50 kd made up of a chondroitin-4-sulfate glycosaminoglycan chain and a 30-kd core protein. NH2-terminal analysis of the core protein showed significant sequence homology with bikunin, a component of the human inter-alpha-trypsin inhibitor (IalphaI) family. A Western blot analysis using anti-human inter-alpha-trypsin inhibitor confirmed the identity of the small chondroitin sulfate proteoglycan as bikunin, and a trypsin inhibitor counterstain assay confirmed its anti-trypsin activity. Examination of serum from patients receiving continuous peritoneal dialysis suggests that free bikunin in peritoneal fluid may be the result of leakage of serum proteins into the peritoneum. Our findings further show that the interaction of serum with peritoneal mesothelial cells offers a new and novel explanation for the presence of bikunin in peritoneal fluid.

Alpha-Globulins↗
PubMed1998

Response of the human peritoneal mesothelial cell to injury: an in vitro model of peritoneal wound healing.

BACKGROUND: The denudation of the peritoneal mesothelium and damage to the underlying interstitium is a frequent finding in patients receiving continuous ambulatory peritoneal dialysis as a treatment for end-stage renal failure. The response of the mesothelium to injury from repeated episodes of infection or from exposure to dialysis fluids has not been extensively studied. The present study describes a simple and reproducible method with which to investigate the response of human mesothelial cells to injury. METHODS: The model of peritoneal injury consists of mechanically wounding a monolayer of human peritoneal mesothelial cells with a glass probe and following the repopulation of the denuded area by time-lapse photomicroscopy. In addition immunohistochemistry was used to follow the response of marker proteins for stress fibers and focal adhesions as well as macromolecules associated with the extracellular matrix. RESULTS: Under serum-free conditions the wound (0.58 +/- 0.094 mm; mean +/- SD; N = 20) closed within 72 +/- 5 hours (N = 8). This rate of healing was enhanced by fetal calf serum, by human serum (10%) and by undiluted spent non-infected dialysate. The repair process over the first 48 hours was the result of cell migration, was independent of cell proliferation and involved the de novo synthesis of several different extracellular matrix components. An early event in the healing process was the rapid reorganization of intracellular stress fibers together with the formation of associated focal adhesions in cells at the wound edge. CONCLUSION: This in vitro model should prove invaluable in characterizing the process of wound healing within the peritoneal cavity, thus allowing a better understanding of the response to infection as well as any effect of dialysis fluids in this pattern of cell behavior.

Blood Physiological Phenomena↗
PubMed1996

IL-1 beta, a major stimulator of hyaluronan synthesis in vitro of human peritoneal mesothelial cells: relevance to peritonitis in CAPD.

The effect of several different growth factors and cytokines on the synthesis of hyaluronan (HA) by human peritoneal mesothelial cells (HPMC) was investigated. Growth arrested HPMC synthesized low levels of HA, but co-culture with PDGF-bb, TGF-beta 1, TNF-alpha, and IL-6 at a concentration of 10 ng/ml all increased HA synthesis between two- to three-fold. At the same concentration IL-1 beta significantly increased the synthesis eight-fold (N = 3; P < 0.05). The effect of IL-1 beta was also dose- and time-dependent and could be totally negated with interleukin-1 receptor antagonist (IL-1 beta RcA). Non-infected and infected dialysate from patients receiving CAPD was also found to stimulate HA synthesis by HPMC. The levels found with non-infected fluid were 4 x 10(4) dpm/ml (N = 6) and 12.9 x 10(4) dpm/ml (N = 6; P < 0.002) and 8.7 x 10(4) dpm/ml (N = 6; P < 0.003) for infected fluid collected one and two days after the commencement of peritonitis. IL-1 beta RcA dramatically reduced the effect of infected but not non-infected dialysate. These results provide new insights into the manner in which HA synthesis is controlled in the mesothelium and suggest that IL-1 beta is a key cytokine in the inflammatory response in CAPD patients.

Adolescent↗
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↗
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↗
PubMed1982

Ascorbic acid absorption in humans: a comparison among several dosage forms.

There have been few studies conducted to determine the efficiency of ascorbic acid absorption in humans. Differences in the extent of its absorption among individuals may contribute to the outcome of clinical trials. Ascorbic acid absorption in four subjects was investigated from several oral dosage forms containing 1 g of the vitamin (solution, tablet, chewable tablet, and timed-release capsule. Approximately 85% of an intravenous dose was recovered in the urine as ascorbic acid and its major metabolites. In contrast, only approximately 30% of the dose was recovered from the solution and tablet forms. A considerably smaller fraction of the dose (approximately 14%) was recovered from the timed-release capsule. There was considerable intersubject variation in ascorbic acid absorption and there appeared to be good and poor absorbers of the vitamin. Consideration should be given to the influence of the extent of ascorbic acid absorption on the results of clinical trails.

Adult↗
PubMed1976

alpha-Chain contacts in the polymerisation of sickle haemogloblin.

Five new double-mutant haemoglobins composed of betaS chains and alpha chains with different substitutions, which are located at the surface of the tetramer, have been prepared. Although all the hybrids are more soluble than deoxyhaemoglobin S, the individual differences between these molecules make it possible to evaluate several regions on the alpha chains for intermolecular contacts in the polymerisation of deoxyhaemoglobin S.

Hemoglobin, Sickle↗