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Biomedical subjects

J M Wang

Publications and source records attributed to J M Wang.

At least 163 records · Page 9Linked to original sources
PubMed1994

Quantitation of endothelial cell specific protein E-9 employing a single monoclonal antibody in an indirect sandwich ELISA.

An indirect enzyme-linked immunosorbent assay is described for the quantitation of protein E-9 which is specifically expressed on human vascular endothelial cells. The assay capitalizes on the dimeric structure of the E-9 protein by utilizing a single monoclonal antibody as both the capture and detection reagent. Detection is achieved by conjugating the Mab with biotin and is followed by the addition of streptavidin peroxidase to provide high sensitivity. Bound activity is measured by enhanced chemiluminescence utilizing standard Amerlite chemistry. The optimised assay is reproducible and is highly sensitive. Using this assay it was possible to detect the presence of E-9 protein in tissue culture media of endothelial cells and in serum samples--in one case even at 1/100 dilution. In vitro, X irradiation resulted in a greater than two-fold increase (P < or = 0.005) in the level of E-9 protein in culture supernatants of human umbilical vein endothelial cells (HUVEC). There are potential applications for measurements of E-9 protein in body fluids and tissue extracts from patients with a vast variety of diseases characterised by vascular endothelial damage and/or activation.

Animals↗
PubMed1994

Oxidation of LDL by nitric oxide and its modification by superoxides in macrophage and cell-free systems.

Oxidative modification of low density lipoproteins (LDL) can increase the atherogenicity of LDL. Here we demonstrate the potential role of nitric oxide (NO) in LDL modification. Dose-dependent alterations of LDL including an increase in electrophoretic mobility, generation of peroxides and degradation of apoprotein B occurred during incubation with NO. Sodium nitroprusside was also shown to increase the electromobility of LDL. Superoxide dismutase (SOD) and catalase did not affect this result. NO treatment also increases the negative charges on other lipoproteins. In addition, we have demonstrated that NO plays a protective role in macrophage or superoxide-induced LDL modification through its neutralizing action.

Animals↗
PubMed1994

Staurosporine restores signaling and inhibits interleukin-8-induced chemotactic desensitization.

Interleukin 8 (IL-8) is a chemotactic cytokine (chemokine) that plays a key role in the accumulation and activation of neutrophils at inflammatory sites. In this report we demonstrate that homologous chemotactic desensitization occurs upon pretreatment of neutrophils with IL-8 or N-formyl-methionyl-leucyl-phenylalanine (FMLP) and results in the inhibition of neutrophil chemotaxis upon subsequent challenge with the same ligand. This homologous chemotactic desensitization could be prevented by pretreating the neutrophils with the protein kinase inhibitor staurosporine, indicating that protein kinases may play an essential role. The attenuation of homologous desensitization by staurosporine restored chemotaxis but was not associated with a change in IL-8 receptor expression, affinity or the rate of ligand internalization, indicating that homologous desensitization does not alter ligand-receptor interaction. Using two-dimensional analysis we have shown that IL-8 induced a rapid serine/threonine phosphorylation of a number of neutrophil substrates the most prominent being phosphoprotein 39 (pp39), extracellular signal-related kinase-1, pp55 and pp66. Prior desensitization of neutrophils with IL-8 blocked all subsequent phosphorylation upon rechallenge with IL-8. However, the desensitization was specific for IL-8 since normal phosphorylation of identical substrates was observed in response to FMLP. When neutrophils were pretreated with staurosporine, prior to desensitization, phosphorylation of pp39 was observed upon restimulation with IL-8. Further study revealed that pp55 and pp66 were not phosphorylated in the presence of staurosporine. Thus, homologous desensitization of neutrophils in response to IL-8 does not result from changes in receptor expression, but rather from a staurosporine-sensitive inactivation of subsequent signal transduction. This desensitization is selective since the cells are able to respond to other ligands.

Alkaloids↗
PubMed1994

Localization of neurokinin A and chromogranin A immunoreactivity in the developing porcine adrenal medulla.

The presence of neurokinin A immunoreactivity was studied in the chromaffin cells of the porcine adrenal medulla and in the nerve fibres innervating the adrenal gland during ontogenic development. For comparison, chromogranin A immunoreactivity was used as a marker for chromaffin cells. Whereas chromogranin A was found in chromaffin cells through all steps in embryonic development, three developmental stages of neurokinin A immunoreactivity could be distinguished. In the first and second trimester of gestation, neurokinin A was observed in some groups of chromaffin cells, but no neurokinin-immunoreactive nerve fibres could be detected. In the last trimester of gestation, neurokinin A-reactive chromaffin cells and nerve fibres were both found in adrenal glands. However, in adrenal glands of neonatal piglets, neurokinin A was found only in nerve fibres and not in chromaffin cells. From these results a hypothesis is proposed that neurokinin A might act as a neurotrophic factor in the early stages of the developing porcine chromaffin cells. Biochemical studies are being performed in order to confirm these morphological results and to study the possible role of neurokinin A as a neurotrophic factor in the adrenal gland.

Adrenal Medulla↗
PubMed1994

Effects of H2O2 on protein tyrosine phosphatase activity in HER14 cells.

Oxidative stress has been implicated in protein phosphorylation and dephosphorylation in cells. In our current studies, H2O2 was shown to reversibly inhibit protein tyrosine phosphatase (PTPase) activity in HER14 cells. H2O2 (150 mM) resulted in 40% inhibition of PTPase activity by 15 min and recovery from inhibition was nearly complete by 60 min. H2O2-induced inhibition or recovery of PTPase activity was not affected by cycloheximide, a protein synthesis inhibitor. L-Buthionine-[S,R]-sulfoximine (BSO), an inhibitor of glutathione synthesis, had no effect on H2O2-induced inhibition of PTPase activity but retarded the recovery of activity. Epidermal growth factor (EGF) and EGTA, a Ca2+ chelator, did not influence H2O2-induced inhibition or recovery of PTPase activity. These results suggest that at least 40% of fibroblast PTPase activity can be regulated by cellular redox activity.

3T3 Cells↗
PubMed1994

Stent of shape-memory alloy for urethral obstruction caused by benign prostatic hyperplasia.

Dilation and positioning a stent in the prostatic urethra have become important alternatives for the management of benign prostatic hyperplasia (BPH), but both have significant drawbacks, namely the need to repeat the treatment in the former case and the conflict between the introducing means and the generation of sufficient expansile force in the latter case. A spiral of a Chinese titanium-nickel alloy with shape memory was implanted in 25 patients with BPH using a self-made coaxial sheath. With a follow-up of 3 to 20 months, the success rate is 92%. There has been no encrustation or migration of the spirals. We deem the spiral of this shape-memory alloy to be a good alternative in patients with BPH who are unfit for surgery.

Aged↗
PubMed1994

Frequency of a deletion polymorphism in the gene for angiotensin converting enzyme is increased in African-Americans with hypertension.

In white populations, a deletion polymorphism in the gene for angiotensin converting enzyme (ACE) appears to be associated with increased risk for myocardial infarction but not for hypertension. In a population-association study in African-Americans, we compared the frequency of the ACE deletion polymorphism in subjects with hypertension versus those with normal blood pressure. The frequency of the ACE deletion allele was greater in African-Americans with hypertension than in those with normal blood pressure (P < 0.05). These findings raise the possibility that in some patient subgroups, sequence variation in or near the ACE gene may contribute to the risk for hypertension.

Adult↗
PubMed1994

Experimental allergic bronchopulmonary aspergillosis in the mouse: immunological and histological features.

C57BL/6 mice treated with intranasal instillation of 100 micrograms of Aspergillus antigen three times a week developed a pulmonary eosinophilia, observed in the bronchoalveolar lavage (BAL) and on histopathological examination. At week 3, the instillation of Aspergillus antigen provoked a 10-fold increase in the BAL cell number and eosinophils were the predominant inflammatory cells (66.4%). Histopathological findings showed focal alveolar lesions with peribronchial and perivascular infiltration of lymphoid cells, numerous eosinophils, epithelioid cells, and granulomas with giant cells. Increases in total IgE and IgG1 levels in BAL fluid (33-fold and 14-fold) and serum (67-fold and 8-fold) were observed also (P < 0.05). IgG1 specific to Aspergillus fumigatus (Af) was detected only in the antigen-treated mice. At 12 weeks, there was a persistent but less intense eosinophilia both in BAL and on histopathological examination accompanied by steadily elevated total IgE and total IgG1 and a higher level of specific IgG1-Af in BAL fluids and sera. No bronchocentric granulomatosis, mucoid impaction nor bronchiectasis could be observed. Data from the study described here showed that in mice repeated exposure to Aspergillus antigen leads to a strong inflammatory pulmonary response, characterized by remarkable pulmonary eosinophilia and elevations of total IgE, total IgG1 and specific IgG1-Af in both BAL and serum, which are the hallmarks of human allergic bronchopulmonary aspergillosis also. However, this inflammation did not induce the chronic histological features of the human disease.

Animals↗
PubMed1994

Genetic contamination of Dahl SS/Jr rats. Impact on studies of salt-sensitive hypertension.

The Dahl salt-sensitive rat (SS/Jr) is a widely used animal model of salt-sensitive hypertension. SS/Jr rats are believed to be highly inbred and uniformly sensitive to the hypertensinogenic effects of sodium chloride, but we have recently observed that SS/Jr rats from Harlan Sprague Dawley, Inc, exhibit considerable variability in their blood pressure response to supplemental dietary salt. To test the possibility that commercially available SS/Jr rats are genetically contaminated and therefore no longer fully inbred, we performed molecular genetic studies and blood pressure measurements in several groups of SS/Jr rats purchased from Harlan Sprague Dawley. We found molecular evidence of heterozygosity and/or atypical allelic variants involving loci on at least five different chromosomes. Many of the rats also failed to exhibit a salt-sensitive blood pressure phenotype. We conclude that SS/Jr rats being sold by the only commercial vendor of Dahl rats in the United States are genetically contaminated and resistant to the hypertensinogenic effects of salt. These findings raise serious questions about the interpretation of research conducted with SS/Jr rats obtained from Harlan Sprague Dawley.

Animals↗
PubMed1994

Role of angiogenesis in patients with cerebral ischemic stroke.

BACKGROUND AND PURPOSE: Stroke is one of the most common causes of mortality and morbidity in the Western world. It results from the occlusion of a cerebral artery followed by severe disturbances in blood supply through microvessels to brain tissue. Despite an extensive literature its pathophysiology is poorly understood, and this has severely impeded the logical development of therapy. METHODS: Brains were obtained from 10 patients aged 46 to 85 years with survival times of 5 to 92 days after their stroke. Infarcted areas and representative control tissues from the contralateral uninvolved brain hemisphere were collected. Microvessel density was measured microscopically. A total of 6520 microvessels were scored in 10,801 areas. The level of activation of the endothelial cells was studied by immunohistochemistry using three monoclonal antibodies, viz, E-9, raised against activated endothelial cells; IG11, recognizing vascular cell adhesion molecule-1; and anti-proliferating cell nuclear antigen. Angiogenic activity in tissue extracts was examined using an in vivo chicken chorioallantoic membrane assay. RESULTS: There was a statistically significant increase in the number of microvessels (Wilcoxon log-rank test; P < or = .01) in 9 of 10 infarcted brain tissues when compared with their contralateral normal hemisphere. In these patients the higher blood vessel counts correlated with longer survival, as ascertained by Spearman's p analysis (P < .02). The number of microvessels filled with blood cells was significantly lower in the infarcted hemispheres (P < .01). In contrast, statistically significant increased numbers of empty microvessels occurred in infarcted tissues compared with the contralateral hemisphere. Monoclonal antibody E-9 reacted weakly with normal-brain vascular endothelial cells; anti-proliferating cell nuclear antigen and IG11 were virtually negative. All three antibodies strongly stained the blood vessels of stroke tissues. The stroke tissues contained angiogenic activity, as shown by the induction of new blood vessels in a chorioallantoic membrane assay. CONCLUSIONS: We have shown that stroke causes active angiogenesis that is more developed in the penumbra. Further experiments are needed to determine if this angiogenesis has beneficial effect.

Aged↗
PubMed1994

VCAM-1 expression on reactive and tumour astrocytes.

In our studies we used a monoclonal antibody recognizing the vascular adhesion molecule (VCAM-1). Tissue samples were collected at autopsy from human brain infarcts and from brain tumours, removed during surgical procedure (Neurological and Neurosurgical Clinic, Cracow). A novel, unexpected finding were VCAM-1-positive fibrous astrocytes in the stroke tissue, and astrocyte-like cells in the tumours. No staining was obtained either in the contralateral hemisphere or outside of ischemic areas. Likewise, no positive staining of cells was seen outside the tumour tissue. The above findings, taken together, strongly support the concept that astrocytes take part in the immune defense during various pathological processes in the human brain.

Astrocytes↗
PubMed1994

[Cephradine delivery into the eye with domestic collagen shield].

The author reports that enhancement by the native made collagen-shield on the permeation of cephradine into the eye is 1.5 times and 1.3 times versus controls when the corneal epithelium is intact or defect, and is 10 times versus frequent eyedrops instillation. On the other hand, the enhancement of the collagen-shield is about the same as that of the soft contact lens impregnated with cephradine solution, but the drug concentration in aqueous humor is increased by collagen-shield 1-2 times than that by the wet soft contact lens. These results indicate that the collagen-shield can effectively increase the drug concentration in aqueous humor.

Animals↗
PubMed1993

Recombinant human interferon-inducible protein 10 is a chemoattractant for human monocytes and T lymphocytes and promotes T cell adhesion to endothelial cells.

The human cytokine interferon-inducible protein 10 (IP-10) is a small glycoprotein secreted by activated T cells, monocytes, endothelial cells, and keratinocytes, and is structurally related to a family of chemotactic cytokines called chemokines. Although this protein is present in sites of delayed-type hypersensitivity reactions and lepromatous leprosy lesions, the biological activity of IP-10 remains unknown. We report here that recombinant human IP-10 stimulated significant in vitro chemotaxis of human peripheral blood monocytes but not neutrophils. Recombinant human IP-10 also stimulated chemotaxis of stimulated, but not unstimulated, human peripheral blood T lymphocytes. Phenotypic analysis of the stimulated T cell population responsive to IP-10 demonstrated that stimulated CD4+ and CD29+ T cells migrated in response to IP-10. This resembles the biological activity of the previously described T cell chemoattractant RANTES. Using an endothelial cell adhesion assay, we demonstrated that stimulated T cells pretreated with optimal doses of IP-10 exhibited a greatly enhanced ability to bind to an interleukin 1-treated endothelial cell monolayer. These results demonstrate that the IP-10 gene encodes for an inflammatory mediator that specifically stimulates the directional migration of T cells and monocytes as well as potentiates T cell adhesion to endothelium.

Antigens, CD↗
PubMed1993

A monoclonal antibody detects heterogeneity in vascular endothelium of tumours and normal tissues.

A new murine monoclonal antibody (MAb), E-9, has been raised using tissue-cultured human umbilical vein endothelial cells. The antigen recognized by this MAb is a peptide of 170 kDa under non-reducing conditions and 96 kDa under reducing conditions. MAb E-9 showed marked heterogeneity in its distribution in various tissues. The antigen recognized by it was present in vascular endothelial cells of all tumours, foetal organs and in regenerating and inflamed tissues. It stained a few normal tissues. However, with the exception of tonsils, staining tended to be weak and limited to a few blood vessels, as revealed by double staining using pan-endothelial antibody (CD31) and antibody to von Willebrand factor, another marker of vascular endothelium. Surprisingly, blood vessels within the placental villi were completely negative. The function of the antigen recognized by MAb E-9 is not known, but its evaluation and use should increase our understanding of angiogenesis.

Adult↗
PubMed1993

Human recombinant macrophage inflammatory protein-1 alpha and -beta and monocyte chemotactic and activating factor utilize common and unique receptors on human monocytes.

The human macrophage inflammatory proteins-1 alpha and -beta (MIP-1 alpha and -beta), which are also known as LD78 and ACT2, respectively, are distinct but highly related members of the chemoattractant cytokine (chemokine) family. rMIP-1 alpha and -beta labeled with 125I specifically bind to human peripheral blood monocytes, the monocytic cell line THP-1, peripheral blood T cells, and the YT cell line. Steady state binding experiments revealed approximately 3000 high affinity binding sites/cell for MIP-1 alpha on human monocytes and on THP-1 cells, with Kd values of 383 pM and 450 pM, respectively. Human MIP-1 alpha and -beta had nearly identical affinities for the binding sites and each competed equally well for binding. Human monocyte chemotactic and activating factor (MCAF), a member of the same chemokine family, consistently displaced about 25% of human MIP-1 alpha and -beta binding on monocytes but not on YT cells, which did not bind MCAF. On the other hand, human rMIP-1 alpha and -beta partially inhibited binding of radiolabeled MCAF to monocytes. Both MIP-1 alpha and -beta were chemotactic for human monocytes. Preincubation of monocytes with human rMIP-1 alpha or -beta markedly reduced cell migration towards the other cytokine, whereas preincubation with human rMCAF only partially desensitized the monocyte chemotaxis response to human rMIP-1 alpha or -beta. These data suggest the existence of three subtypes of receptors, i.e., one unique receptor shared by MIP-1 alpha and -beta, a second unique receptor for MCAF, and a third species that recognizes both MCAF and MIP-1 peptides.

Binding Sites↗
PubMed1993

Identification of RANTES receptors on human monocytic cells: competition for binding and desensitization by homologous chemotactic cytokines.

RANTES (regulated on activation, normal T expressed and secreted) is a member of the chemotactic cytokine (chemokine) beta subfamily. High affinity receptors for RANTES have been identified on a human monocytic leukemia cell line THP-1, which responded to RANTES in chemotaxis and calcium mobilization assays. Steady-state binding data analyses revealed approximately 700 binding sites/cell on THP-1 cells with a Kd value of 400 pM, comparable to that expressed on human peripheral blood monocytes. The RANTES binding to monocytic cells was competed for by monocyte chemotactic and activating factor (MCAF) and macrophage inflammatory protein 1 (MIP-1) alpha, two other chemokine beta cytokines. Although MCAF and MIP-1 alpha competed for RANTES binding to monocytes with apparent lower affinity (with estimated Kd of 6 and 1.6, nM respectively) both of these cytokines effectively desensitized the calcium mobilization induced by RANTES. The chemotactic response of THP-1 cells to RANTES was also markedly inhibited by preincubation with MCAF or MIP-1 alpha. In contrast, RANTES did not desensitize the THP-1 calcium mobilization and chemotaxis in response to MCAF or MIP-1 alpha. These results, together with our previous observations that RANTES did not compete for MCAF or MIP-1 alpha binding on monocytic cells, indicate the expression of promiscuous receptors on monocytes that recognize one or more cytokines within the chemokine beta family.

Calcium↗