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Biomedical subjects

J Shen

Publications and source records attributed to J Shen.

At least 397 records · Page 22Linked to original sources
PubMed1995

Pharmacologically induced regression of chronic transplant rejection.

Chronic rejection, characterized by a progressive obliterative arteriopathy, is a major cause of graft failure in long-surviving human transplants for which there is no effective treatment. Leflunomide, an isoxazol derivative, has been shown to be a novel immunomodulatory drug that profoundly suppresses the immune response. In this study, 58 Fisher-344 rats received cardiac transplantation from Lewis rats. All the recipients were given CsA at 2.5 mg/kg for 5 days postoperatively. Without further treatments, the arterial intima was progressively injured by mononuclear cell infiltration and Ab deposition. Smooth muscle cell and fibroblast proliferation in the intima became a predominant phenomenon by day 90. CsA was ineffective in controlling the progress of arterial intimal thickening when treatment began on day 30. Leflunomide at 5 mg/kg failed to control arterial intimal thickening by day 60 when therapy began on day 30. However, the progress of arterial intimal thickening was significantly inhibited by day 90 when the dosage of leflunomide had been increased to 10 mg/kg on day 60. Combined therapy with leflunomide and CsA at 5 mg/kg for 30 days dramatically reversed the arterial thickening by day 60. After increasing the dosages of both leflunomide and CsA to 10 mg/kg on day 60, the combination therapy steadily controlled the chronic rejection. Only the combination therapy significantly down-regulated circulating antidonor IgM and IgG titers. In rat smooth muscle cell culture, this same drug combination had a synergistic inhibitory effect on proliferation. Therefore, the combination therapy of leflunomide and CsA could reverse and control the progress of chronic rejection, while leflunomide, at higher dosage as a monotherapy, could stabilize chronic rejection in this model. The mechanism of the regression of chronic rejection by leflunomide and cyclosporine may be related to their in vitro abilities to control not only lymphocyte but smooth muscle cell proliferation, as well. The synergistic effect of these two drugs on vascular smooth muscle cell proliferation in vitro may be an important part of this novel activity. This unique feature holds intriguing possibilities for treating established chronic rejection.

Animals↗
PubMed1995

A comparison of the conformation of sucrose octasulfate, free and bound to acidic fibroblast growth factor.

The conformation of sucrose octasulfate free in solution has been determined based on high-resolution NMR spectroscopy. Three-bond 1H-1H scalar coupling constants, laboratory and rotating frame NOEs, long-range 1H-13C scalar coupling constants, and chemical-shift temperature and ionic-strength dependence were used, aided by molecular mechanics calculations. By modification of a pulse sequence designed for measuring long-range 1H-13C coupling constants, it was possible to obtain an accurate value for the 3JC2f-H1g despite the confounding presence of 3JHH of similar value. Free sucrose octasulfate appears to assume a conformation significantly different from any of the eight conformations observed bound to acidic fibroblast growth factor, as determined in a previous X-ray crystallographic study [X. Zhu, B.T. Hsu, and D.C. Rees, Structure, 1 (1993) 27-34]. Strong electrostatic interactions between guest and host may be the dominant factor in deformation of sucrose octasulfate. The implications of this study for protein-carbohydrate interactions and the effects of the presence of sulfate groups on the flexibility of sucrose are also discussed.

Animals↗
PubMed1995

Early induction of interferon-independent virus-specific ICAM-1 (CD54) expression by flavivirus in quiescent but not proliferating fibroblasts--implications for virus-host interactions.

West Nile Virus (WNV) infection of human embryonic fibroblasts can induce expression of ICAM-1 by two distinct mechanisms. An early and direct mechanism occurs within 2 hr of virus infection which is cytokine independent, and an indirect mechanism occurs within 24 hr of virus infection and is regulated by the release of IFN-type 1. Virus-inactivated, conditioned supernatants removed from WNV-infected fibroblast cultures at 4 hr did not alter ICAM-1 expression on fresh, untreated fibroblasts, whereas conditioned supernatants from 24-hr-infected cultures induced small increases in ICAM-1 expression after incubation for 24 hr but not after 4 hr. These studies also demonstrate that the expression of ICAM-1 on fibroblasts in response to flavivirus is cell-cycle dependent. WNV can only induce increased ICAM-1 expression in quiescent fibroblasts in G0 phase. In contrast, induction of ICAM-1 after exposure to types 1 and 2 IFN is not cell-cycle dependent. Other viruses, including double-stranded DNA viruses, vaccinia, and adenovirus 2 and 5 and the single, positive-stranded RNA alphavirus, Semiliki Forest virus, did not induce ICAM-1 expression on fibroblasts after 24 hr. Another alphavirus, Ross river, was able to induce ICAM-1 but only by the indirect mechanism of type 1 IFN-dependent release. The closely related flavivirus, Kunjin, induced increased ICAM-1 expression in a manner similar to WNV. The ability of flaviviruses to induce increased ICAM-1 expression directly within a few hours of infection may be an important virus-host survival strategy promoting cell-cell adhesion and hence possible further viral infection/replication.

Adenoviruses, Human↗
PubMed1995

Exon skipping by overexpression of a Drosophila heterogeneous nuclear ribonucleoprotein in vivo.

Heterogeneous nuclear ribonucleoproteins (hnRNPs) are abundant RNA-binding proteins that are implicated in splicing regulation. Here we investigate the role of a Drosophila hnRNP in splicing regulation in living animals. We find that overexpression of the Drosophila hnRNP HRB98DE leads to skipping of all internal exons in the Drosophila dopa decarboxylase (Ddc) pre-mRNA in vivo. These results indicate that HRB98DE has a splicing activity that promotes use of terminal splice sites. The effect of excess HRB98DE on Ddc splicing is transient, even though high levels of HRB98DE persist for at least 24 hr. This suggests that Drosophila larvae can induce a compensating mechanism to counteract the effects of excess HRB98DE.

Animals↗
PubMed1995

SCN5A mutations associated with an inherited cardiac arrhythmia, long QT syndrome.

Long QT syndrome (LQT) is an inherited disorder that causes sudden death from cardiac arrhythmias, specifically torsade de pointes and ventricular fibrillation. We previously mapped three LQT loci: LQT1 on chromosome 11p15.5, LQT2 on 7q35-36, and LQT3 on 3p21-24. Here we report genetic linkage between LQT3 and polymorphisms within SCN5A, the cardiac sodium channel gene. Single strand conformation polymorphism and DNA sequence analyses reveal identical intragenic deletions of SCN5A in affected members of two unrelated LQT families. The deleted sequences reside in a region that is important for channel inactivation. These data suggest that mutations in SCN5A cause chromosome 3-linked LQT and indicate a likely cellular mechanism for this disorder.

Amino Acid Sequence↗
PubMed1995

Prognosis of patients > or = 70 years of age with non-Q-wave acute myocardial infarction compared with younger patients with similar infarcts and with patients > or = 70 years of age with Q-wave acute myocardial infarction.

In this study, 70 patients > or = 70 years of age admitted to the coronary care unit with non-Q-wave acute myocardial infarction (AMI) were followed prospectively for 1 year, and the clinical course in these patients was compared with that in 61 patients < 70 years with non-Q-wave AMI and 56 patients > or = 70 years with Q-wave AMI. Compared with the younger patients with non-Q-wave AMI, older patients were more likely to develop atrial fibrillation (23% vs 8%; p < 0.05) and congestive heart failure (53% vs 30%; p < 0.01), and less likely to receive thrombolytic therapy (9% vs 28%; p < 0.01), cardiac catheterization (41% vs 72%; p < 0.01), and coronary angioplasty (20% vs 39%; p < 0.05). Hospital mortality did not differ significantly between older and younger non-Q-wave AMI patients (10% vs 3%), but 1-year mortality was higher in the elderly (36% vs 16%; p = 0.02). Elderly patients with Q-wave AMI had more in-hospital complications, including death (25% vs 10%; p < 0.05), than elderly patients with non-Q-wave AMI. In contrast, postdischarge mortality was higher in elderly patients with non-Q-wave AMI, so that total mortality at 1 year was similar in the 2 groups. Overall, elderly patients with non-Q-wave AMI accounted for 62% of all deaths occurring during the first year after discharge (relative risk 2.6 compared with other groups; p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗
PubMed1995

Dielectric relaxation spectroscopy and some applications in the pharmaceutical sciences.

With a few exceptions, dielectric relaxation spectroscopy (DRS) has been largely neglected by pharmaceutical scientists, despite the potential for this technique as a noninvasive and rapid method for the structural characterization and quality control of pharmaceutical materials. DRS determines both the magnitude and time dependency of electrical polarization (i.e. the separation of localized charge distributions) by either measuring the ability of the material to pass alternating current (frequency domain DRS) or by investigating the current that flows on application of a step voltage (time domain DRS). DRS is thus (i) sensitive to molecular mobility and structure, (ii) non-invasive, and (iii) employs only mild stresses (a weak electromagnetic field) in order to measure the sample properties. The technique covers a broad-band frequency window (from 10(-5) to 10(11) Hz) and therefore enables the investigation of a diverse range of processes, from slow and hindered macromolecular vibrations and restricted charge transfer processes (such as proton conductivity in nearly dry systems) to the relatively fast reorientations of small molecules or side chain groups. The dielectric response provides information on (i) structural characteristics of polymers, gels, proteins, and emulsions, (ii) the interfacial properties of molecular films, (iii) membrane properties, (iv) water content and states of water (and the effects of water as a plasticizer), and (v) lyophilization of biomolecules. This review article details the basis of dielectric theory and the principles of measuring dielectric properties (including a comprehensive account of measurement artifacts), and gives some applications of DRS to the pharmaceutical sciences.

Chemistry, Pharmaceutical↗
PubMed1995

Binding of phosphorus-containing inhibitors to thermolysin studied by the Poisson-Boltzmann method.

Zinc endopeptidase thermolysin can be inhibited by a series of phosphorus-containing peptide analogues, Cbz-Gly-psi (PO2)-X-Leu-Y-R (ZGp(X)L(y)R), where X = NH, O, or CH2; Y = NH or O; R = Leu, Ala, Gly, Phe, H, or CH3. The affinity correlation as well as an X-ray crystallography study suggest that these inhibitors bind to thermolysin in an identical mode. In this work, we calculate the electrostatic binding free energies for a series of 13 phosphorus-containing inhibitors with modifications at X, Y, and R moieties using finite difference solution to the Poisson-Boltzmann equation. A method has been developed to include the solvation entropy changes due to binding different ligands to a macromolecule. We demonstrate that the electrostatic energy and empirically derived solvation entropy can account for most of the binding energy differences in this series. By analyzing the binding contribution from individual residues, we show that the energy of a hydrogen bond is not confined to the donor and acceptor. In particular, the positive charges on Zn and Arg 203, which are not the acceptors, contribute significantly to the hydrogen bonds between two amides of ZGpLL and the thermolysin.

Amino Acid Sequence↗
PubMed1995

Conservative and nonconservative mutations in proteins: anomalous mutations in a transport receptor analyzed by free energy and quantum chemical calculations.

Experimental studies on a bacterial sulfate receptor have indicated anomalous relative binding affinities for the mutations Ser130-->Cys,Ser130-->Gly, and Ser130-->Ala. The loss of affinity for sulfate in the former mutation was previously attributed to a greater steric effect on the part of the Cys side chain relative to the Ser side chain, whereas the relatively small loss of binding affinity for the latter two mutations was attributed to the loss of a single hydrogen bond. In this report we present quantum chemical and statistical thermodynamic studies of these mutations. Qualitative results from these studies indicate that for the Ser130-->Cys mutation the large decrease in binding affinity is in part caused by steric effects, but also significantly by the differential work required to polarize the Cys thiol group relative to the Ser hydroxyl group. The Gly mutant cobinds a water molecule in the same location as the Ser side chain resulting in a relatively small decrease in binding affinity. Results for the Ala mutant are in disagreement with experimental results but are likely to be limited by insufficient sampling of configuration space due to physical constraints applied during the simulation.

Bacterial Proteins↗
PubMed1995

Expression of adhesion molecules in cultured human pulmonary microvascular endothelial cells.

HPMECs were successfully isolated by differential trypsinization from peripheral lung lobes. The cells proliferated rapidly in EGM-MV with 10% FBS and were serially cultivated for more than 20 passages (1:4 split ratio) in vitro. Cells were characterized as endothelial based upon their cobblestone morphology, the presence of factor VIII-related antigen, incorporation of DiI-Ac-LDL, tubule-like structure formation in Matrigel, and positive staining for ACE. Adhesion molecules were tested at passage 3 and passage 12. Cells demonstrated intense staining for PECAM-1 both unstimulated and stimulated with TNF-alpha (20 ng/ml). The adhesion molecules ICAM-1, VCAM-1, ELAM-1, and P-selectin differed in expression on unstimulated cells. ICAM-1 was constitutively expressed on unstimulated cells and the expression was increased by TNF-alpha stimulation (20 hr). In contrast, VCAM-1, ELAM-1, and P-selectin were not detected on unstimulated cells but were detected after stimulation with TNF-alpha. The inducibility of adhesion molecules was different. VCAM-1 (10 hr) and ELAM-1 (4 hr) were expressed more strongly than P-selectin (minutes to 4 hr). The adhesion molecule profile found on passage 12 was the same as on passage 3. CD36 was not detected on both unstimulated and stimulated (4 and 8 hr) cells. The peak of adhesion of HL-60 cells to TNF-alpha activated HPMEC monolayers was around 8 hr. The results indicate that HPMEC can be continuously grown in vitro for many passages without losing their adhesion molecule expression. This expression of adhesion molecules confirms that HPMECs might be a good in vitro model in the understanding of various aspects of pulmonary microvascular endothelial cell function and may be useful as the basis for studies of adhesion molecule targeted therapies of pulmonary inflammatory diseases.

Cell Adhesion Molecules↗
PubMed1995

Rural development and rural to urban migration in China 1978-1990.

"Economic reforms since the late 1970s have brought about significant changes in rural China. A large number of surplus rural labourers have been released from the agricultural sector and there has been a massive transition of rural residents from agricultural to non-agricultural employment. These changes will be analyzed by examining the changes in the employment structure of rural residents.... It is found that town and country populations tended to move to towns at the intra-provincial level, but to cities at the interprovincial level. Out-migrants from cities tended to move to cities at both the intra- and inter-provincial levels."

Agriculture↗
PubMed1995

Inactivation of Kupffer cells after prolonged donor fasting improves viability of transplanted hepatic allografts.

Data from recent studies suggest that donor fasting imparts a beneficial effect on the viability of transplanted hepatic allografts. Because starvation may temporarily inactivate Kupffer cells, and because these cells are the likely mediators of liver injury after prolonged preservation-reperfusion, the purpose of this study is to establish a link between improved organ viability and Kupffer cell inactivation caused by donor allograft fasting. In an in vivo rat liver transplant model, 48 hours of donor fasting (1) improved allograft viability, (2) significantly decreased Kupffer cell phagocytosis, and (3) significantly decreased cytokine (tumor necrosis factor [TNF]) production postrevascularization. These data validate work from previous studies demonstrating that donor fasting improves allograft viability and furthermore support our previous research implicating activation of Kupffer cells as a causative agent of cold ischemia-preservation injury.

Animals↗
PubMed1995

Cardiac sodium channel mutations in patients with long QT syndrome, an inherited cardiac arrhythmia.

Long QT syndrome (LQT) is an inherited cardiac disorder that causes syncope, seizures and sudden death from ventricular tachyarrhythmias. We used single-strand conformation polymorphism (SSCP) and DNA sequence analyses to identify mutations in the cardiac sodium channel gene, SCN5A, in affected members of four LQT families. These mutations include two identical intragenic deletions and two missense mutations. These data suggest that SCN5A mutations cause LQT. The location and character of these mutations suggest that this form of LQT results from a delay in cardiac sodium channel fast inactivation or altered voltage-dependence of inactivation.

Amino Acid Sequence↗
PubMed1995

Transgenic rabbits with the integrated human 15-lipoxygenase gene driven by a lysozyme promoter: macrophage-specific expression and variable positional specificity of the transgenic enzyme.

15-Lipoxygenase is expressed in foamy macrophages of atherosclerotic lesions and has been implicated in the oxidative modification of low density lipoprotein during early stages of atherogenesis. To establish an animal model of 15-lipoxygenase overexpression, we created transgenic rabbits that express at high level the human 15-lipoxygenase in monocyte-derived macrophages but not in liver, heart, kidney, lung, or other tissues. The expression level of the enzyme in monocyte-derived macrophages is comparable to that of interleukin 4 (IL4)-treated human monocytes, but more than 20-fold higher than in macrophages of normal rabbits. The transgenic enzyme oxygenates linoleic acid to 13S-hydroperoxy-9, 11 (Z,E)-octadecadienoic acid (13-HODE), and arachidonic acid to a mixture of 12S-hydroperoxy-5, 8, 10, 14 (Z,Z,E,Z)-eicosatetraenoic acid (12S-HETE), and 15S-hydroperoxy-5, 8, 11, 14 (Z,Z,Z,E)-eicosatetraenoic acid (15S-HETE). The 12-HETE/15-HETE ratio varied between 0.3 and 5.4, indicating a remarkable variability in the positional specificity of the transgenic enzyme. Macrophages from normal rabbits consistently produced 12S-HETE as the major oxygenation product. 15-Lipoxygenase-overexpressing rabbits may be used for further mechanistic studies on the implication of lipoxygenase in atherogenesis; they are also an ideal model for testing the in vivo action of 15-lipoxygenase inhibitors.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗