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Biomedical subjects

M Cheng

Publications and source records attributed to M Cheng.

At least 19 recordsLinked to original sources

Evidence for a Ras-dependent extracellular signal-regulated protein kinase (ERK) cascade.

The small GTP-binding protein Ras appears to be required for transformation and differentiation induced by tyrosine kinases. The Ras requirement may be limited to a few tyrosine kinase-regulated signaling pathways or may be universal for all tyrosine kinase actions. Because both Ras and the microtubule-associated protein 2 kinases ERK1 and ERK2 have been implicated in events that lead to neurite outgrowth, we explored the possibility that Ras and ERKs may lie on the same signaling pathway. Utilizing PC-12 rat adrenal pheochromocytoma cell lines that contain a dominant inhibitory Ras mutant (S17N-Ras(H)), we found that Ras was required for stimulation of the ERK cascade by nerve growth factor but apparently not by the heterotrimeric G protein activator AlF4-. Within this cascade, Ras appears to be upstream of an ERK activator, raising the intriguing possibility that Ras may directly regulate a serine/threonine protein kinase.

Aluminum

Human soluble IL-6 receptor: its detection and enhanced release by HIV infection.

By using a fluorescence sandwich ELISA for the quantification of soluble human IL-6R, normal human PBMC were found to be induced to release IL-6R into culture supernatant by stimulation with PHA. Furthermore, certain promonocyte cell lines and human T-cell leukemia virus I (HTLV-I)-positive cell lines produced sIL-6R into culture supernatants constitutively. However, this was not found with HTLV-I negative T cell lines and Burkitt's B cell line. In addition, generation of supernatant IL-6R of the promonocyte cell line was significantly increased 27-fold after PMA treatment and sevenfold after infection with HIV. The released IL-6R molecules were characterized as an apparent m.w. of 50 to 55 kDa by both size-exclusion HPLC and immunoprecipitation of the soluble protein with IL-6R-specific mAb followed by SDS-PAGE analysis. Furthermore, increased levels of serum IL-6R were detected in blood donors seropositive for HIV. Moreover, the released IL-6R could bind efficiently to purified rIL-6 on solid phase and suppressed the proliferative responses of PBMC. These results suggest that the release of soluble IL-6R might be linked to regulatory functions of immune responses induced by IL-6 stimulation during normal and human retrovirus-infected cell growth and differentiation.

Enzyme-Linked Immunosorbent Assay

Cytoskeletal modulation of plasma membrane events induced by interferon-alpha.

Cytochalasin B, a drug that alters microfilament structure, was found to modulate interferon-alpha (IFN-alpha)-induced changes in ion fluxes, in motional freedom of spin probes, and lateral diffusion of surface antigens. These changes occur in Daudi cells inherently sensitive to the antiproliferative signal of IFN-alpha, but not in insensitive cells, and were associated with the antiproliferative signal previously. The biophysical effects of cytochalasin B were detected by flow cytometric quantitation of membrane potential using an oxonol dye, by electron spin resonance (ESR) spectrometry, and by measurements of fluorescence recovery after photobleaching (FRAP) of surface antigens using a laser-interactive cell imaging system. Cytochalasin B treatment increased an IFN-alpha-induced membrane potential shift by -5 mV. The motional freedom of 5-doxyl-stearic acid changed from 0.67 to 0.63, as expressed by the order parameter, S, with IFN-alpha treatment and was prevented by cytochalasin B. Changes in the lateral diffusion of surface antigens induced by IFN-alpha treatment, D = 5.3 x 10(-10) without treatment and D = 7.8 x 10(-10) cm2/s with treatment, was blocked by cytochalasin B. In contrast, the microtubule stabilizers taxol and D2O and the microtubule depolymerizing colcemid were ineffective at dose levels sufficient to cause the characteristic cell physiological alterations of these agents. These results implicate microfilaments but not the microtubule system in transduction of the antiproliferative signal by IFN-alpha in Daudi cells.

Actin Cytoskeleton

Soluble interleukin-6 receptor is released from receptor-bearing cell lines in vitro.

Soluble interleukin-6 receptor (sIL-6R) was found to be spontaneously released from human myeloma cell line U266 cells into culture supernatant, and was quantitatively measured with a fluorescence sandwich enzyme-linked immunosorbent assay employing antibodies specific to IL-6R. The supernatant IL-6R was generated only from IL-6R-positive cell lines; myeloma cell lines RPMI8226 and PRMI1788, and myelomonocytic cell lines U937, THP-1, and HL-60. In contrast, it was not released from the IL-6R-negative cells; T cell line Molt-4 and Burkitt lymphoma cell line Raji. SDS-PAGE analysis of the soluble IL-6R from U266 cells suggested a molecular weight of approximately 50-55 kDa, 25-30 kDa smaller than the mature cell surface receptor. These results suggest that the generation of soluble IL-6R may be a maker of myeloma cells and myelomonocytic cells.

Cell Line

Effects of vitamin A on retinal pigment epithelial cells in vitro.

Human retinal pigment epithelial (RPE) cell monolayers were used in an in vitro wound-healing experiment. Wound closure were studied over time in the presence of various concentrations of all-trans retinol (vitamin A). It was found that 5-20 micrograms/ml (17.5-70 mumol/l) of vitamin A significantly inhibited wound closure by inhibiting cell migration and proliferation. In a collagen gel-contraction model, similar doses of vitamin A partially inhibited cell contractility. Cellular morphology, as assessed by phase-contrast microscopy, changed from a regular polygonal shape to an elongated stellate shape. Indirect immunofluorescence patterns of fibronectin, actin, tubulin, and vimentin were altered after exposure to vitamin A. All these effects were reversible after removal of the drug from the medium. Modulation of RPE growth and cell-mediated contraction may be useful in the treatment of ocular proliferative diseases.

Cell Count

Glycolipid receptor binding specificity of exoenzyme S from Pseudomonas aeruginosa.

By use of the tlc overlay procedure we have shown that exoenzyme S extracted from cultures of Pseudomonas aeruginosa specifically binds to the glycolipids asialoGM1, asialoGM2 and to a lesser extent lactosyl ceramide. More significantly, strong binding was also observed to the glycerolipid receptor we have detected for Helicobacter pylori (Lancet ii, 238-241.1989). Exoenzyme S can be extracted in a toxic and nontoxic form. Toxicity correlated with ability to bind the H. pylori receptor. This species was the only receptor detected in the most sensitive cell lines. The relative binding of exoenzyme S to the ganglio series glycolipids and the glycerolipid receptor was modified in a reciprocal manner in the presence of metal ions, suggesting that exoenzyme S has two interrelated receptor binding sites.

ADP Ribose Transferases

Mitochondrial protein import.

A dynamic picture of the mitochondrial protein import pathway is emerging, with conformational alteration a critical feature both preceding and following membrane translocation. The mediators of these steps of conformational alteration, as well as steps of recognition, translocation, and proteolytic cleavage, appear to be proteins. Using powerful tools of genetics and biochemistry, in years to come it should be possible to determine the precise molecular function of these proteins in mediating these novel reactions.

Animals

The surgical anatomy of the sinoatrial node.

The sinoatrial nodes (SAN) were observed, dissected, and measured on 95 adults and 30 child hearts under a dissection microscope. The majority of the SANs in adults are characterized by their pale color, firm consistency, and the location in relation to the penetration of the SAN artery, and they can be located in the superior part of the terminal sulcus. The SANs in children, however, are not easily discerned. The variation of the apex of the right auricular crest and the notch in the superior part of the terminal sulcus have been described, and the present authors suggested that the trigone of the SAN could be used as an important landmark to identify the SAN. The surface features on the SAN, its relationship to the surrounding myocardium and its surgical significance during operation are further discussed.

Adult

Toward a unified theory of sexual mixing and pair formation.

Sexually transmitted diseases such as gonorrhea, syphilis, herpes, and AIDS are driven and maintained in populations by epidemiological and sociological factors that are not completely understood. One such factor is the way in which people mix sexually. In this paper, we outline a unified approach to modeling sexual mixing structures, where such structures are defined in terms of a set of axioms for a finite number of distinct groups of people. Theorems for homosexual, heterosexual, and arbitrary group mixing are presented, leading to a representation of all mixing structures defined by the axioms. The representation and its parameters are interpreted in terms of intergroup affinities for sexual mixing. The use of the approach in sexually transmitted disease modeling is discussed.

Female

[Effect of processing of toxic constituents in radix Phytolaceae].

This paper reports the effect of different processing technologies on the main toxic constituents in Radix Phytolaceae using its contents of phytolaccatoxin and histamine as index. The study was carried out to probe into the processing principle and improve the processing technology of Radix Phytolaceae. The result is gratifying.

Drugs, Chinese Herbal

Biomechanical properties and collagen fiber orientation of temporomandibular joint discs in dogs: 2. Tensile mechanical properties of the discs.

The tensile properties of TMJ discs in dogs were investigated using soft-tissue tensile and stress-relaxation tests. The data obtained suggested that (1) the mediolateral tensile stiffness (199.7 and 234.0 MPa), tensile strength (46.73 and 69.99 MPa), and equilibrium moduli (30.0 and 30.1 MPa) of anterior and posterior bands of the disc, respectively, were much greater than those of the middle part (46.6, 14.71, and 18.4 MPa, respectively); (2) the difference in fracture strain of various parts was not significant; (3) the physiologic strain region of the discs was about 4%; and (4) the properties of low stress/relaxation rate and nonlinear relationship of the instantaneous elastic response to various strains may represent lower permeability of TMJ discs.

Analysis of Variance

Effect of lateral mobility of fluorescent probes in lipid mixing assays of cell fusion.

Monolayers of human erythrocytes, immobilized on a cover slip, were induced to fuse by polyethylene glycol (mol wt 8,000). The mobility of fluorescent probes, 1-oleoyl-2-[12-[(7-nitro-2,1,3-benzoxadizol-4-yl)amino]dodecanoyl] phosphatidyl-choline (C12-NBD-PC), from labeled cells to unlabeled cells was monitored by video-enhanced fluorescence microscopy. A dequenching curve was obtained from the measurement of fluorescence intensities of pairs of fused cells over time. The dequenching curve and the curve obtained from macroscopic measurements of a cell monolayer (described in the preceding article) were compared and discussed. The slow probe transfer rate between a pair of fused cells was explained by a diffusion model based on membrane area conservation and the geometry of the fusion lumen. An equivalent lumen between two fused cells, thought to be the main rate limitation of probe mobility after fusion, was calculated to be approximately 130 nm in diameter. Lumens of 75 nm in diameter were observed by electron microscopy. Thus, the rate of macroscopic fluorescence dequenching depends not only upon the fusion efficiency, but also upon the number of simultaneous fusion partners, the geometry of their contact points, and the lateral mobility of the fluorescent probes through these points. The relative fusion efficiency can be derived only from the saturation dequenching values.

4-Chloro-7-nitrobenzofurazan

[Content of histoamine in processed Phytolacca americana L].

The contents of histamine in processed Phytolacca americana has been determined by colorimetry. The result shows that the contents tend to decrease by 23.3% when the herb is shredded, but get a bit higher again when fried with vinegar. The method may be useful in the quality control of the herb.

Colorimetry

Total and direct-reacting bilirubin values by automated methods compared with liquid chromatography and with manual methods for determining delta bilirubin.

This study compares total and direct-reacting bilirubin values in 40 serum samples from patients with various diagnoses, as measured by automated methods (Beckman Synchron CX-5, Beckman Astra 8, Kodak Ektachem 700) and HPLC and by a manual method for delta bilirubin. For total bilirubin, within-run CVs were less than 6%. The Ektachem 700 method underestimated bilirubin with serum samples from patients with Crigler-Najjar syndrome and from newborns in whom unconjugated bilirubin concentrations were increased but conjugated bilirubins were not present or were present only in small amounts. The Astra 8 and Synchron CX-5 methods were inaccurate with cholestatic serum samples, in which conjugated bilirubin concentrations were increased and other compounds such as bile acids could be expected to interfere. We conclude that each automated method examined provides reasonable estimates for total and direct-reacting bilirubin values for routine clinical use. The need for each laboratory to select the appropriate bilirubin method for its particular situation is obvious.

Autoanalysis

Selection of therapeutic agents for intraocular proliferative disease 3. Effects of fluoropyrimidines on cell-mediated contraction of human fibroblasts.

We evaluated the efficacy of 5-fluorouracil (5-FU) and 5-fluorouridine (5-FUR) as anticontractile agents in an in vitro model that measures the ability of human and rabbit fibroblasts to contract a collagen gel. Human fibroblasts were more contractile than rabbit fibroblasts, required higher concentrations of 5-FUR for inhibition of contraction, and were able to tolerate higher doses of the drug. 5-FU failed to inhibit contraction of human fibroblasts at concentrations up to 1.5 mM (200 micrograms ml-1), but significantly inhibited rabbit fibroblast contraction, although to a lesser degree than 5-FUR under similar conditions. 5-Fluorouridine (100 microM, 26 micrograms ml-1) inhibited contraction of human fibroblasts by more than 80%, whereas only 10 microM (2.6 micrograms ml-1) 5-FUR was required for 90% inhibition of rabbit fibroblast contraction. 5-FUR was cytotoxic to rabbit fibroblasts at concentrations of 200 microM (52 micrograms ml-1) or greater, but 800 microM was non-toxic to human fibroblasts after 96 hr exposure. At concentrations of 1.5 mM, 5-FU had no significant effects on the viability of either human or rabbit fibroblasts. These results suggest that 5-FUR may prove useful for the treatment of human ocular proliferative disorders in which contraction of fibrocellular elements is thought to play a part. They also suggest that fluoropyrimidine dosages found to be effective in a rabbit model of proliferative vitreoretinopathy may be less effective in humans and that 5-FUR levels shown to be toxic to rabbit-derived cells may be tolerated by humans cells.

Cells, Cultured

Oxygenation during chest physiotherapy of very-low-birth-weight infants: relations among fraction of inspired oxygen levels, number of hand ventilations, and transcutaneous oxygen pressure.

Chest physiotherapy (CPT) to clear the airway of infants requiring assisted ventilation is a routine procedure in neonatal intensive care units. This study was done to describe the relationships between fraction of inspired oxygen (FiO2) levels and the number of hand ventilations (independent variables) and variations of transcutaneous oxygen pressure (TcPO2) from baseline among intubated very-low-birth-weight (VLBW) infants receiving CPT. The study consisted of 30 observations of 13 intubated VLBW infants receiving CPT. Data were obtained by observations during routine CPT procedures performed by the nurses in charge of the infants. Stepwise multiple regression showed that hand ventilations and FiO2 levels significantly accounted for the variability in TcPO2 levels during CPT. Pearson correlation analysis showed that there was a negative correlation between FiO2 and TcPO2 and between the number of hand ventilations and TcPO2.

Blood Gas Monitoring, Transcutaneous