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Biomedical subjects

H S Yu

Publications and source records attributed to H S Yu.

At least 91 records · Page 5Linked to original sources

Induction of Ca2+ oscillations by vasopressin in the presence of tetraethylammonium chloride in cultured vascular smooth muscle cells.

The change of cytosolic Ca2+ concentration ([Ca2+]i) caused by vasopressin was examined in indo-1-loaded A7r5 smooth muscle cells by use of the high-performance laser cytometer and ratiometric fluorescence method. Vasopressin (100 nM) caused an initial rapid rise and a delayed increase in [Ca2+]i (n = 6). However, in the presence of tetraethylammonium chloride (10 mM), vasopressin consistently triggered sustained Ca2+ oscillations which were preceded by a large peak of [Ca2+]i. The latency for the development of this huge increase in [Ca2+]i prior to the occurrence of sustained Ca2+ oscillations was always the same. The frequency and amplitude of this type of Ca2+ oscillation varied depending upon the extracellular Ca2+ concentration. Ca(2+)-free solution did not completely suppress the sustained Ca2+ oscillations, but caffeine (20 mM) effectively abolished them. The present findings indicate that in A7r5 smooth muscle cells, the sustained Ca2+ oscillations triggered by vasopressin in the presence of tetraethylammonium chloride were mainly due to Ca2+ release from IP3-sensitive Ca2+ stores and Ca2+ influx from extracellular space, and did not require the pacemaker activity derived from the surface membrane. Moreover, the vasopressin-induced change in [Ca2+]i appeared to be linked to pertussis toxin-insensitive GTP-binding protein(s).

Animals↗

Defective spontaneous and bacterial lipopolysaccharide-stimulated production of interleukin-1 receptor antagonist by polymorphonuclear neutrophils of patients with active systemic lupus erythematosus.

Interleukin-1 receptor antagonist (IL-1ra) binds competitively to IL-1 receptors but does not transduce the signal which blocks the biological activities induced by IL-1. In this study, polymorphonuclear neutrophils (PMN) and mononuclear cells (MNC) from the patients with active systemic lupus erythematosus (SLE) (n = 11), inactive SLE (n = 13) and normal individuals (n = 13) were compared for the IL-1ra producing capacity of these cells. PMN and MNC at a concentration of 1 x 10(6) cells/ml were incubated with medium alone (spontaneous) or stimulated with lipopolysaccharide (LPS, 100 ng/ml) for 24 h. The IL-1ra concentration in the supernatants was quantified by ELISA method. Both spontaneous and LPS-stimulated production of IL-1ra by PMN, but not by MNC, of active SLE were significantly lower than that of inactive SLE or normal groups. Prednisolone (1 and 5 micrograms/ml) did not change the production of IL-1ra by normal PMN either spontaneously or LPS-stimulation in in vitro study. Moreover, the IL-1ra producing capacity of PMN in seven active SLE on admission and after intensive immunosuppressive treatment was measured. These results suggest that the defective IL-1ra production by SLE-PMN is relevant to disease activity and may be regarded as a new indicator of disease activity in patients with active SLE.

Adolescent↗

Abnormal splenic and thymic IL-4 and TNF-alpha expression in MRL-lpr/lpr mice.

The MRL-lpr/lpr and MRL-(++) mice were studied for the expression of cytokines in the spleen, lymph node, thymus, kidney and brain through the reverse transcription-polymerase chain reaction (RT-PCR). The frequencies of IL-4 and TNF-alpha expression in the thymus and spleen were significantly higher in MRL-lpr/lpr mice than in MRL-(++) mice from the age of 17 to 32 weeks. More importantly, IL-4 transcript was demonstrated in the early rather than in the terminal stage of the lupus disease. At the 20th week, MRL-lpr/lpr mice with active disease exhibited higher concentrations of IL-1 alpha, IL-6 and TNF-alpha in serum than MRL-(++) mice. Interestingly, in MRL-lpr/lpr but not MRL-(++) mice, the IL-6 concentration in cultured supernatants of the thymic cells was significantly higher than that of the splenic or lymph node cells. On the other hand, IL-6 and IL-1 beta were expressed in the brain and kidney of MRL-lpr/lpr mice but not of MRL-(++) mice. Cultured MRL-lpr/lpr mesangial cells could also express IL-6 but to a lesser extent. These results suggest that the abnormal splenic and thymic IL-4 and TNF-alpha expression may predispose the development of autoimmune reactions. The expression of IL-1 beta and IL-6 in the brain and kidney may be implicated in the damage of these two organs in MRL-lpr/lpr mice.

Animals↗

Production of the third component of complement (C3) by peripheral polymorphonuclear neutrophils of the patients with rheumatoid arthritis.

Normal human polymorphonuclear neutrophils (PMN) can spontaneously produce the third component of complement (C3) in in vitro culture as detected by ELISA. This C3-producing capacity of PMN can be augmented by TNF-alpha (20 ng/ml) and bacterial lipopolysaccharide (100 ng/ml), but not by IL-1 beta or IL-8. The C3 production by PMN was found to be temperature dependent and was suppressed by the addition of protein inhibitor. The C3 mRNA in PMN could be detected by reverse transcription assisted polymerase chain reaction (RT-PCR) after TNF-alpha or LPS stimulation for 6 hours. To further understand C3 production by peripheral blood PMN in rheumatoid arthritis (RA), spontaneous and TNF-alpha stimulated production of C3 by peripheral PMN were compared in 15 cases of active RA, 15 inactive RA and 15 normal individuals. We failed to find any significant difference among the three groups. We conclude that PMN plays a negligible role in C3 hypercomplementemia in patients with active RA.

Arthritis, Rheumatoid↗

Responses of cutaneous microcirculation to cold exposure and neuropathy in vibration-induced white finger.

In order to clarify the mechanisms of cold temperature on vibration-induced white finger (VWF), three groups of forestry chain saw operators served as the subjects of this study. Groups I and II worked in a cold high-altitude area, and group III worked in a warm low mountain climate. Group I workers had VWF sign [VWF(+)] but groups II and III did not [VWF(-)]. In the present study, finger nailfold microcirculation before and after cold exposure, finger skin sympathetic alpha receptor response, and sympathetic skin response were measured. The results showed that there was no significantly different neuropathy in any of the groups. However, vibration-induced microcirculatory disturbances including blood stasis and red blood cell aggregation were more prominent in VWF(+) group, whereas both of the VWF(-) groups (groups II and III) had no significant difference in microcirculatory disturbances. This suggests that the severity of the microcirculatory disturbances is dose-effect correlated and is aggravated by cold in vibration syndrome. Thus it can be realized that a cold working environment is an important contributing factor in causing microcirculatory disturbances of VWF.

Altitude↗

Vibration induced neurophysiological and electron microscopical changes in rat peripheral nerves.

This study was conducted to clarify the effects of vibration on the peripheral nerves. Rat tails were exposed to vibration (acceleration 56.9 m/s2, frequency 60 Hz, amplitude 0.4 mm for two or four hours daily, six days a week. The maximum motor conduction velocity (MCV), the amplitude of evoked response, and the motor distal latency were measured on rat tail nerves every two months. Thin sections of tail nerves were examined under the electron microscope after 200, 500, and 800 hours of vibration. Neurophysiological and ultrastructural changes in tail nerves increased with the dose of vibration. In the groups exposed to vibration the MCVs were significantly reduced after a vibration time up to 400 hours, whereas the motor distal latency was not delayed significantly until 600 vibration hours. The ultrastructural changes were (1) detachment of the myelin sheath from the axolemma, (2) constriction of the axon, (3) protrusion of the myelin sheath into the axon, (4) accumulation of vacuoles in paranodal regions, and (5) dilatation of the Schmidt-Lanterman incisures. The ultrastructural changes induced by vibration in the paranodal regions and myelin sheaths were possibly responsible for the reduction in MCVs.

Animals↗

Cerebrospinal fluid interleukin-6, prostaglandin E2 and autoantibodies in patients with neuropsychiatric systemic lupus erythematosus and central nervous system infections.

Cerebrospinal fluid (CSF) from patients with a variety of central nervous system (CNS) disorders was assayed for cytokines, prostaglandins, and autoantibodies. CSF interleukin-6 (IL-6) in patients with CNS infection (374.24 +/- 92.61 pg/mL) and neuropsychiatric systemic lupus erythematosus (NP-SLE) (71.40 +/- 5.89 pg/mL) were significantly higher than in patients with CNS inflammation (33.92 +/- 29.36 pg/mL) or controls (non-inflammatory CNS diseases) (4.35 +/- 3.00 pg/mL). Interleukin-1 beta, interferon alpha, and tumor necrosis factor alpha were undetectable in these samples: CSF prostaglandin E2 (PGE2) also exhibited similar patterns as IL-6. CSF immunoglobulin G (IgG) in patients with NP-SLE (8.84 +/- 1.80 mg/dL) was much higher than in patients with CNS infection (4.65 +/- 3.09 mg/dL), CNS inflammation (2.54 +/- 1.24 mg/dL), or controls (2.11 +/- 1.03 mg/dL). CSF autoantibodies against calf thymus antigens were present in patients with NP-SLE but not in patients with CNS infection as demonstrated by immunoblot. These results suggest that high IL-6 and PGE2 in CSF favors the diagnosis of CNS infection, while modestly elevated IL-6, high IgG, and autoantibodies against calf thymus antigens in CSF are the features of NP-SLE.

Autoantibodies↗

Increased production of tumor necrosis factor-alpha and release of soluble CD4 and CD8 molecules, but decreased responsiveness to phytohemagglutinin in patients with nasopharyngeal carcinoma.

Twelve patients with nasopharyngeal carcinoma (NPC) were studied for spontaneous and phytohemagglutin (PHA)-stimulated production of cytokines, soluble markers and [3H] thymidine incorporation by mononuclear cells. The same number of patients with skin cancer and healthy subjects were used as control groups. Our results showed that the NPC group produced much more tumor necrosis factor-alpha (TNF alpha), soluble CD4 (sCD4) and CD8 (sCD8) in PHA-stimulated mononuclear cell supernatants than those in the other two groups. The levels of soluble interleukin-2 receptor (sIL-2R) and gamma-interferon (IFN gamma) in PHA-stimulated supernatants were at the same high level in the NPC and healthy subjects groups while the concentrations were much lower in the skin cancer group. We also noticed that the early stage group in NPC patients had higher levels of interleukin-1 alpha (IL-1 alpha), TNF alpha, IFN gamma and sIL-2R in both spontaneous and PHA-stimulated mononuclear cell supernatants. The stimulation index of PHA-responsiveness was 155, 5.2 and 37, respectively, in the healthy subjects, skin cancer and NPC groups. The PHA-responsiveness was depressed in both the NPC and skin cancer groups. It seems that cancer patients have an impaired T cell mitogenic response after mitogen stimulation. NPC patients had better immune response than skin cancer patients in immune factor release or PHA-responsiveness.

Adolescent↗

Establishment and characterization of a continuous human basal cell carcinoma cell line from facial skin (I) cytological behavior of early passages.

Basal cell carcinomas (BCCs) are one of the most common skin malignant diseases of human beings in the course of a lifetime. BCC is a slow growing tumor of epithelial origin. Most of the BCC tumors have a normal diploid DNA configuration and have been successfully passaged for a short-term only. We now report a long-term culture (BCC-1/KMC) of human BCC derived from the undifferentiated type of facial BCC tumor on the thermal traumatic scar, which was aneuploidy and subculture for more than 100 passages. The population doubling time of BCC-1/KMC cells at the third passage was 38.86 hours. This immortalized and tumorigenic cell line expresses epithelial markers of keratin filaments and desmosomes. The genetic markers of this cell line have HLA-A2, A24, B27, B35, Cw3, DR2 and DR12. The BCC-1/KMC cells have successfully adapted to grow in a cheaper commercial medium (RPMI 1640) and at a moderate concentration of calcium (0.4 mM).

Animals↗

[Vitamin A status in patients with cholelithiasis].

In order to assess the status of vitamin A and determine the frequency of vitamin A deficiency in adults with cholelithiasis, we measured plasma vitamin A levels in 40 healthy subjects and 64 patients with cholelithiasis. Among them, 40 were icteric and the remaining 24 were anicteric. Vitamin A levels were determined in biopsied liver tissues in 26 patients with cholelithiasis. Plasma levels of vitamin A in patients with cholelithiasis was significantly lower than that of healthy subjects (P < 0.05). A significant difference of vitamin A in liver tissue was noted between these two patient groups (P < 0.05). The frequency of vitamin A deficiency, defined as a plasma vitamin a level below 90 micrograms/dl, was 50% (32/64) in cholelithiasis patients, 65% (26/40) in jaundiced patients, and 25% (6/24) in jaundice-free patients. We conclude that biochemical deficiency of vitamin A occurs in patients with cholelithiasis. We suggest that vitamin A should be evaluated in patients with cholelithiasis, and an effective therapy should be initiated to prevent vitamin A deficiency.

Adult↗

Changes of lipid peroxide levels in blood and liver tissue of patients with obstructive jaundice.

Plasma lipid peroxide levels, hereafter referred to as PLP levels, were measured in a group of 40 apparently healthy controls and 64 cholelithiasis patients, 40 with and 24 without jaundice. Hepatic lipid peroxide (HLP) levels were also measured in 26 patients, 15 with and 11 without jaundice. There was a significantly higher mean concentration of PLP in the jaundiced patients than in the control or jaundice-free cases. However, the difference in PLP levels between the jaundice-free and the control cases was insignificant. Meanwhile, patients with jaundice had significantly higher HLP levels than those without jaundice. In the jaundiced cases, the increased PLP and HLP levels were clearly related to the serum levels of bilirubin respectively. In addition, the HLP levels were positively correlated with the PLP levels; however, in the non-jaundiced cases, there was little evidence of these two relationships. Patients with or without jaundice had lower plasma vitamin E levels in comparison to the control cases. The correlation of plasma vitamin E and PLP levels was weak in all of the jaundiced. However, when we subdivided the jaundiced into two groups, the correlation was strong in those with plasma vitamin E levels < 8.5 micrograms/ml, while the correlation was weak in those with plasma vitamin E levels > 8.5 micrograms/ml. Consequently, these results suggest that there is an involvement of lipid peroxidation in liver cells damaged by obstructive jaundice in cholelithiasis patients and there exists a negative correlation between low vitamin E and lipid peroxide levels in plasma.

Adult↗

Prostaglandin E2 suppresses phytohemagglutinin-induced immune responses of normal human mononuclear cells by decreasing intracellular glutathione generation, but not due to increased DNA strand breaks or apoptosis.

Prostaglandin E2 (PGE2) at concentrations more than 1 x 10(-8) M markedly suppressed the cell proliferation and release of soluble molecules of interleukin-2 receptor (sIL-2R), CD4 (sCD4) and CD8 (sCD8) from phytohemagglutinin (PHA)-stimulated normal human mononuclear cells (MNC) in a dose-related manner. To further elucidate the subcellular mechanism of the inhibitory effect of PGE2 on PHA-stimulated MNC, intracellular concentration of glutathione (GSH) in PHA-stimulated MNC was sequentially measured from day 1 to day 3 by enzymic method. Furthermore, the effect of PGE2 on nuclear DNA including DNA strand breaks in alkali treatment and DNA fragmentation (apoptosis) of PHA-stimulated MNC were also measured. We found intracellular GSH levels were significantly decreased in the early stage of lymphocyte activation (day 1), but no evidence of increased DNA strand breaks or apoptotic process appeared in 3-day culture. In addition, butathione sulfoximine (a specific GSH inhibitor) and dibutyryl cyclic AMP also exhibited both proliferation inhibition and GSH-decreasing effects on PHA-stimulated MNC as well as PGE2. These results suggest that the immunosuppressive effect of PGE2 is mediated by the decreased generation of intracellular GSH, but not by the increased DNA strand breaks or apoptotic mechanism in the cells.

Antimetabolites↗

Melatonin inhibits the proliferation of retinal pigment epithelial (RPE) cells in vitro.

The possible antiproliferative effect of melatonin on retinal pigment epithelial (RPE) cells in vitro was investigated. Bovine RPE cells cultured in Ham's F12 medium supplemented with 10% fetal bovine serum had a nuclear density of 73.6 +/- 6.1 nuclei/mm2 at 72 h after seeding. The nuclear density at this time-point was doubled if either 50 or 100 ng/ml human epidermal growth factors (hEGF) was added to the culture medium. When these hEGF-stimulated cells were treated with melatonin from 10 to 500 pg/ml, the proliferation was suppressed with a dose-response relationship. At 250 and 500 pg/ml melatonin, the nuclear densities of the melatonin-treated cells were similar to those of the control cells. Using mitotically active SV-40 transformed human fetal RPE cells cultured in a serum-free medium, melatonin was also shown to be antiproliferative. In the presence of 500 pg/ml melatonin, the proliferation of these cells was inhibited to 77% as compared to the control. These results were further supported by the reduced [H3]thymidine uptake in the melatonin-treated cells. We propose that melatonin, at physiologic concentrations, has an antiproliferative effect, and that cultured RPE cells stimulated to proliferate by either hEGF treatment or SV-40 transfection are responsive to melatonin. Melatonin may either inhibit mitosis in actively dividing cells or modulate hEGF action.

Animals↗

Coexistence and relationship of antikeratinocyte and antimelanocyte antibodies in patients with non-segmental-type vitiligo.

To test for autoantibodies in patients with vitiligo, skin biopsies from 16 patients with active vitiligo and 12 patients with stable vitiligo were examined by direct immunofluorescence. In periodate-lysine-paraformaldehyde-fixed biopsy specimens, the presence of IgG deposits in keratinocytes and the number of keratinocytes with focal IgG in active vitiligo were significantly greater than in stable vitiligo. To test whether the antibodies to normal human keratinocytes or melanocytes are present in vitiligo, we used an indirect immunofluorescent test and enzyme-linked immunosorbent assay to test the serum of 43 patients. With unfixed viable melanocytes, we found a granular pattern of IgG staining on the plasma membrane of melanocytes incubated with patients' sera but not in cells incubated with the control sera. With methanol-fixed melanocytes, however, we found a homogeneous pattern of IgG staining in the cytoplasm of melanocytes. With unfixed viable keratinocytes as targets, there was no deposit of IgG on the cells. A homogeneous pattern of IgG binding in the cytoplasm of methanol-fixed keratinocytes suggested the presence of antikeratinocyte autoantibodies to cytoplasmic keratinocyte components. The fluorescence staining for IgG binding was more prominent in active or extensive vitiligo. Vitiligo sera were cytotoxic for melanocytes but not for keratinocytes in vitro. Antimelanocytic antibodies may play a role in melanocytotoxicity, whereas antikeratinocyte antibodies may occur secondary to cellular damage.

Antibody-Dependent Cell Cytotoxicity↗

Progressive alterations of cytokeratin expressions in the process of chronic arsenism.

Recent studies of an endemic occurrence of chronic arsenism in a limited area on the southwest coast of Taiwan are focusing on its cytokeratin analysis in hopes of tracing the disease's biochemical expression. Specimens were obtained from uninvolved skin and arsenical cancers including Bowen's disease, basal cell carcinoma, and squamous cell carcinoma. In this study, we used two-dimensional polyacrylamide gel electrophoresis to analyse cytokeratin expression. Progressive alterations in cytokeratin expression were found in various skin lesions. These include an expression of K16 in the uninvolved skin; K16 and K6 in Bowen's disease; and K16, K6 and K17 in squamous cell carcinoma and basal cell carcinoma. In addition, we found that the K1 isoelectric variants shifted to more acidic forms with the complete absence of K1 in basal cell carcinoma. K16 expression in uninvolved skin indicates that it is nevertheless in a hyperproliferative status. K17 was expressed in squamous cell carcinoma and basal cell carcinoma, but not in Bowen's disease. The progressive impairment of phosphorylation of K1 and K2 in the process of chronic arsenism provides us with a suitable model for studying the biological significance of phosphorylation in intermediate filaments during chemical carcinogenesis.

Arsenic Poisoning↗

Sodium-dependent ascorbic and dehydroascorbic acid uptake by SV-40-transformed retinal pigment epithelial cells.

The present data confirmed previous studies with other cell types that ascorbic acid and dehydroascorbic acid are transported through different transporters into SV-40-transformed retinal pigment epithelial cells. These experiments were performed on cells grown on 96-well culture plates. Ascorbic acid was taken up into the cell by a high-affinity transporter with Km = 0.041 mmol/l and a low Vmax of 2.74 pmol/min/well. Dehydroascorbic acid was taken up by a low-affinity transporter with Km = 5.67 mmol/l; however, the Vmax was 325.5 pmol/min/well. The uptake of both ascorbic acid and dehydroascorbic acid was dependent on the sodium concentration. The uptake of ascorbic acid does not involve oxidation-reaction steps because the uptake of [14C]-ascorbate was unaffected by the presence of an excess amount of unlabelled dehydroascorbic acid.

Ascorbic Acid↗

Characterization of functional antagonism of adenosine and isoproterenol in guinea-pig atrial myocytes.

The role of adenosine action in modulating the action potential duration (APD) of single isolated guinea-pig atrial myocytes during beta-adrenergic stimulation with isoproterenol was investigated. Membrane potential and ionic currents were measured by the tight-seal, whole-cell patch-clamp technique. Adenosine (10 microM) antagonized isoproterenol (20 nM)-stimulated L-type calcium inward current (ICa). In the presence of isoproterenol, adenosine (10 microM) also increased holding and late outward currents. Isoproterenol (20 nM) caused a significant prolongation in atrial action potential at the level of 0 mV, rather than at 90% repolarization. Adenosine shortened atrial action potential dose-dependently in the presence of isoproterenol. The respective values for EC50 and maximally shortened action potential duration at 0 mV of adenosine with or without isoproterenol (20 nM) were 3.0 vs. 2.2 microM and 42.9 vs. 30.4 ms. In summary, during beta-adrenergic stimulation with isoproterenol, in guinea-pig atrial myocytes, adenosine elicited the specific K(+)-outward current and concomitantly attenuated the isoproterenol-stimulated ICa. These phenomena may account for the modulation of atrial action potential by adenosine with or without isoproterenol effect.

Action Potentials↗

Prevalence of shoulder and upper-limb disorders among workers in the fish-processing industry.

A cross-sectional study was conducted among fish-processing workers to evaluate the prevalence of shoulder and upper-limb discomforts and to assess the associated ergonomic risk factors. A prestructured interview, a medical check-up, and job analyses were performed to determine musculoskeletal disorders among 207 workers in eight factories. The results showed shoulder girdle pain (30.9%), epicondylitis (14.5%), and carpal tunnel syndrome (15.0%) as the three most common soft-tissue disorders. The odds ratio of shoulder girdle pain was 1.6 (95% CI 1.1-2.5) among the workers who performed tasks with repetitive movement of their upper limbs, while it was 1.8 (95% CI 1.2-2.5) for the workers who sustained forceful movement of their upper limbs during work. Women taking oral contraceptives had a 2.0 times higher odds ratio for carpal tunnel syndrome than did other women. It would appear that untrained or unskilled workers were prone to suffer from musculoskeletal disorders.

Adult↗