Biomedical subjects

D Zhang

Publications and source records attributed to D Zhang.

At least 199 records · Page 11Linked to original sources

PubMed2000

Eradication of murine mammary adenocarcinoma through HSVtk expression directed by the glucose-starvation inducible grp78 promoter.

Gene therapy strategies employing the HSVtk/ganciclovir (GCV) suicide gene offer promising approaches towards the treatment of metastatic breast cancer. These include bystander effects on non-transduced tumor cells, lower systemic toxicity, and the possibility of inducing immunity against the tumor. Previously we have demonstrated the ability of the grp78 stress-inducible promoter to stimulate expression of reporter genes within the tumor microenvironment. However, experimental evidence demonstrating the ability of this promoter to activate therapeutic agents within the breast cancer environment causing tumor eradication is needed prior to clinical trials. In this report, we test the efficacy of the grp78 promoter in a retroviral system to drive the expression of the HSVtk suicide gene in a murine mammary adenocarcinoma cell line (TSA) in syngeneic, immune-competent hosts. Our results show that under glucose-starvation conditions in vitro, the expression of HSVtk and GCV induced cell death are enhanced in tumor cells in which the HSVtk gene is driven by the internal grp78 promoter compared to cells in which the Moloney murine leukemia virus LTR drives HSVtk. In in vivo studies, in tumors in which the HSVtk gene is driven by the grp78 promoter, GCV treatment causes complete tumor eradication, whereas tumors persist when the HSVtk gene is driven by the retroviral LTR. Our study suggests that the grp78 promoter may be useful to enhance the effectivity of therapeutic agents within a breast tumor. In addition, it is shown that immune memory is induced in syngeneic, immune-competent hosts. This new retroviral vector might therefore be useful for breast cancer gene therapy.

Adenocarcinoma↗

PubMed2000

TAK1 is activated in the myocardium after pressure overload and is sufficient to provoke heart failure in transgenic mice.

The transforming-growth-factor-beta-activated kinase TAK1 is a member of the mitogen-activated protein kinase kinase kinase family, which couples extracellular stimuli to gene transcription. The in vivo function of TAK1 is not understood. Here, we investigated the potential involvement of TAK1 in cardiac hypertrophy. In adult mouse myocardium, TAK1 kinase activity was upregulated 7 days after aortic banding, a mechanical load that induces hypertrophy and expression of transforming growth factor beta. An activating mutation of TAK1 expressed in myocardium of transgenic mice was sufficient to produce p38 mitogen-activated protein kinase phosphorylation in vivo, cardiac hypertrophy, interstitial fibrosis, severe myocardial dysfunction, 'fetal' gene induction, apoptosis and early lethality. Thus, TAK1 activity is induced as a delayed response to mechanical stress, and can suffice to elicit myocardial hypertrophy and fulminant heart failure.

Activating Transcription Factor 6↗

PubMed2000

A novel immunoassay for the evaluation of rod outer segment digestion in cultured retinal pigment epithelial cells.

In this work a novel enzyme-linked immunoabsorbent assay quantifying residual rod outer segments in the medium of rod outer segment-challenged retinal pigment epithelial cells is described. A retinal pigment epithelial cell line (D407) that produces low level of cathepsin D, and a primary human retinal pigment epithelial cell culture (HRPE51) that has normal cathepsin D levels, were challenged with bovine rod outer segments. At 3 days post-challenge, the amount of undigested or residual bovine rod outer segments left in the culture medium was quantified by an enzyme-linked immunoabsorbent assay. An antibody raised against bovine rod outer segments, which had been purified and labelled with nitroiodophenyl haptens, was used in the assay. The sensitivity of the immunoassay was less than 10(2) bovine rod outer segments per mL and the signal followed a linear curve, saturating around 10(6) bovine rod outer segments per mL. HRPE51 cells had no residual bovine rod outer segments present in the medium following a challenge with 10(4) bovine rod outer segments per mL. In the medium of D407 cells, residual bovine rod outer segment levels were higher at all bovine rod outer segment concentrations when compared to the residual bovine rod outer segment levels in HRPE51 cells, suggesting that D407 cells have a lower digestive capacity. These results demonstrated that the immunoassay for detecting bovine rod outer segments is a sensitive and reliable technique that can be used to quantify the amount of residual bovine rod outer segments, following bovine rod outer segment challenge of retinal pigment epithelial cells.

Animals↗

PubMed2000

Evidence for coassembly of mutant GABAC rho1 with GABAA gamma2S, glycine alpha1 and glycine alpha2 receptor subunits in vitro.

Functional coassembly of gamma-aminobutyric acid (GABA)C rho1 subunits with GABAA (alpha1, beta2, and gamma2S) or glycine (alpha1, alpha2, and beta) subunits was examined using two-electrode voltage-clamp recordings in the Xenopus laevis oocyte expression system. To facilitate this study, we took advantage of the unique gating and pharmacological properties of two mutant rho1 subunits, rho1(T314A) and rho1(T314A/L317A). When the rho1(T314A) subunit was coexpressed with GABA gamma2S, glycine alpha1 or glycine alpha2 subunits, GABA response properties were different from those of homomeric rho1(T314A) receptors. Additionally, the sensitivity of heteromeric rho1(T314A) and gamma2S receptors to picrotoxinin (PTX) blockade of GABA-evoked responses was altered compared to that of homomeric rho1(T314A) receptors. Changes in GABA response properties and picrotoxinin sensitivity were also observed when rho1(T314A) subunits were coexpressed with wild-type rho1 subunits. When rho1(T314A/L317A) subunits were coexpressed with GABA gamma2S, glycine alpha1 or glycine alpha2 subunits, suppression by GABA of spontaneously active current was reduced compared to that of homomeric rho1(T314A/L317A) receptors. Recovery of the spontaneous current from inhibition by GABA for GABA rho1(T314A/L317A)/gamma2S heteromeric receptors displayed an additional component. Coinjection of wild-type rho1 with gamma2S cRNAs at a ratio of 1 : 1 resulted in a > 10-fold reduction in GABA-evoked current. Furthermore, coexpression of wild-type rho1 and gamma2S subunits was found to shift the GABA dose-response curve. Our results provide functional evidence that the GABAC rho1 subunit can coassemble with the GABAA gamma2S subunit, and, at least in its mutated form, rho1 can also form heteromeric receptors with glycine alpha1 or alpha2 subunits in vitro.

Animals↗

PubMed2000

Expression of GABA(C) receptor rho1 and rho2 subunits during development of the mouse retina.

Retinal gamma-aminobutyric acid type C (GABA(C)) receptors consist of rho subunits. Here we report our results from a competitive PCR and patch-clamp electrophysiology study quantifying rho subunit message and characterizing GABA(C) receptor-mediated currents at different stages of mouse retinal development. Mouse rho1 message is first detected at postnatal day 6 (P6), increases significantly until P9 and remains at this level through adulthood, whereas mouse rho2 message does not appear until P9, peaks at P15 and remains at this level through adulthood. There is an approximate twofold excess of rho1 compared to rho2 message at most stages of development, which persists in adulthood. Functional GABA(C) receptors are detected in acutely dissociated bipolar cells of P9 or older mouse retina. Early in development (P9-10), GABA(C) receptors are composed solely of rho1 subunits, but subsequently contain rho1 and/or rho2 subunits (by P11 and later). These findings are intriguing because the onset and rapid increase in rho subunit transcription and functional expression match the initiation and active period of bipolar cell differentiation in retinal development as well as the stage of eye opening and initial visual experience in the rodent. The investigation of mouse rho subunits here forms a basis for future studies on the role of GABA(C) receptors in retinal development.

Age Factors↗

PubMed2000

Vascularized plantaris tendon graft: anatomic study of the donor.

In order to determine new vascularized donor tendons for grafting, a detailed anatomic study of the plantaris tendon and vascular connections with the posterior tibial artery or its branches was undertaken in 20 legs of 10 fresh adult cadavers. A histologic evaluation was also undertaken. Findings demonstrated that there is a close vascular connection between the crural fascial-linked section of the plantaris tendon and the posterior tibial artery. Through its rich surrounding paratenon, the blood supply of the tendon is provided by two to four transfascial branches of the posterior artery in the lower-middle portion of the leg. Out of these branches, one or two anastomosing arteries (more than 1.0 mm in diameter), together with accompanying veins, consistently emerge 5 to 8 cm from the insertion of the plantaris tendon. Histologic observations demonstrate the reliable vascularity of the paratenon and crural fascia. The authors consider this vascularized tendon donor a good option for grafting. A composite tendofascial flap with vascularized pedicle from the posterior tibial artery or an isolated vascularized tendon graft from its branches would both be alternative techniques for clinical vascularized tendon grafting.

Adult↗

PubMed2000

Genotoxic markers among butadiene polymer workers in China.

While 1,3-butadiene is carcinogenic in rodents, cancer causation in humans is less certain. We examined a spectrum of genotoxic outcomes in 41 butadiene polymer production workers and 38 non-exposed controls, in China, to explore the role of butadiene in human carcinogenesis. Because in vitro studies suggest that genetic polymorphisms in glutathione S-transferase enzymes influence genotoxic effects of butadiene, we also related genotoxicity to genetic polymorphisms in GSTT1 and GSTM1. Among butadiene-exposed workers, median air exposure was 2 p.p.m. (6 h time-weighted average), due largely to intermittent high level exposures. Compared with unexposed subjects, butadiene-exposed workers had greater levels of hemoglobin N-(2,3,4-trihydroxybutyl)valine (THBVal) adducts (P < 0.0001) and adduct levels tended to correlate, among butadiene-exposed workers, with air measures (P = 0.03). Butadiene-exposed workers did not differ, however, from unexposed workers with respect to frequency of uninduced or diepoxybutane-induced sister chromatid exchanges, aneuploidy as measured by fluorescence in situ hybridization of chromosomes 1, 7, 8 and 12, glycophorin A variants or lymphocyte hprt somatic mutation. Also among the exposed, greater THBVal levels were not associated with increases in uninduced sister chromatid exchanges, aneuploidy, glycophorin A or hprt mutations. Butadiene-exposed workers had greater lymphocyte (P = 0.002) and platelet counts (P = 0.07) and lymphocytes as a percentage of white blood cells were moderately correlated with greater THBVal levels (Spearman's phi = 0.32, P = 0.07). Among butadiene-exposed workers, neither GSTM1 nor GSTT1 genotype status predicted urinary mercapturic acid butanediol formation, THBVal adducts, uninduced sister chromatid exchanges, aneuploidy or mutations in the glycophorin A or hprt genes. Overall, the study demonstrated exposure to butadiene in these workers, by a variety of short-term and long-term measures, but did not show specific genotoxic effects, at the chromosomal or gene levels, related to that exposure.

Adult↗

PubMed2000

Horn fly (Diptera: Muscidae) saliva targets thrombin action in hemostasis.

The horn fly, Hematobia irritans (L.), is an important pest of livestock because the adult stage of both sexes are aggressive blood-feeders. Remarkably, even though horn fly adults feed recurrently on their hosts as ectoparasites, these flies lack the ADP-responsive antiplatelet aggregation and vasodilatory antihemostatic systems described for other blood-feeding Diptera. Horn fly salivary gland extracts do interfere with the normal coagulation process as demonstrated by the recalcification time assay. Using this as a baseline, the effects of saliva on recalcification time, activated partial thromboplastin time, prothrombin time, and thrombin time were measured to determine which arm(s) of the coagulation cascade might be impacted. Factor-deficient plasma assays also were used to measure possible perturbations in clotting. Gland-free saliva delayed the recalcification time as well as the activated partial thromboplastin time, prothrombin time, and thrombin time. Saliva also further delayed clotting times of plasmas deficient in factor V, factor VIII, and factor XIII, indicating that other factors in the coagulation cascade were inhibited. Although horn fly saliva did not alter the ability of deficient plasma reconstituted with factor X to clot, it did inhibit deficient plasma reconstituted with factor II (thrombin). Antithrombin activity in saliva was confirmed by its ability to interfere with thrombin hydrolysis of fibrinogen, its normal substrate, and by its inhibition of thrombin action on a chromagenic substrate that mimics the hydrolytic site of fibrinogen. Thus, horn fly saliva contains a factor that specifically targets thrombin, a key component in the coagulation cascade. While the biochemical mechanisms of inhibition may vary, this antihemostatic characteristic is shared with other zoophilic Diptera such as black flies, Simulium spp., and tsetse, Glossina morsitans morsitans Westwood, that feed on ungulates.

Animals↗

PubMed2000

Accuracy and completeness of Medicare claims data for surgical treatment of breast cancer.

BACKGROUND: Although a number of studies have used Medicare claims data to study trends and variations in breast cancer treatment, the accuracy and completeness of information on surgical treatment for breast cancer in the Medicare data have not been validated. OBJECTIVES: This study assessed the accuracy and completeness of Medicare claims data for breast cancer surgery to determine whether Medicare claims can serve as a source of data to augment information collected by cancer registries. METHODS: We used the Surveillance, Epidemiology and End Results (SEER) Cancer Registry-Medicare data and compared Medicare claims on surgery with the surgery recorded by the SEER registries for 23,709 women diagnosed with breast cancer at > or =65 years of age from 1991 through 1993. RESULTS: More than 95% of women having mastectomies according to the Medicare data were confirmed by SEER. For breast-conserving surgery, 91% of cases were confirmed by SEER. The Medicare physician services claims and inpatient claims were approximately equal in accuracy on type of surgery. The Medicare outpatient claims were less accurate for breast-conserving surgery. In terms of completeness, when the 3 claims sources were combined, 94% of patients receiving breast cancer surgery according to SEER were identified by Medicare. CONCLUSIONS: The combined Medicare claims database, which includes the inpatient, outpatient, and physician service claims, provides valid information on surgical treatment among women known to have breast cancer. The claims are a rich source of data to augment the information collected by tumor registries and provide information that can be used to follow long-term outcomes of Medicare beneficiaries.

Breast Neoplasms↗

PubMed2000

Antiretroviral use and pharmacy-based measurement of adherence in postpartum HIV-infected women.

BACKGROUND: Antiretroviral treatment for HIV-infected women is standard during pregnancy to prevent vertical transmission, but data on postpartum therapy for the mother are lacking. OBJECTIVE: The objective of this study was to examine the impact of provider and patient characteristics on receipt of antiretroviral therapy and pharmacy-based measurement of adherence by postpartum HIV-infected women. RESEARCH DESIGN: This was a retrospective cohort study. SUBJECTS: The study included 2,648 New York State Medicaid-enrolled HIV-infected women who delivered from January 1993 through October 1996 and were followed up through September 1997. MEASURES: From Medicaid claims in the first postpartum year, the study examined any prescribed antiretroviral therapy and, among women treated >2 months, adherence, defined as > or =80% days covered by prescribed therapy from first to last antiretroviral prescription. RESULTS: Antiretroviral therapy was prescribed for 681 (26%) study women. Of 292 women treated >2 months, 28% were adherent on the basis of the pharmacy-based measure. The proportion of treated women was highest in 1996 (40%), and adherence was best in 1995 (44%) when most women took monotherapy. The adjusted odds ratios (AORs) of treatment were 1.67 (95% CI, 1.24 to 2.25) for women receiving HIV-focused services and 2.71 (95% CI, 1.99 to 3.69) for women with a provider in an HIV-related specialty. The AORs of adherence were greater for women with HIV-focused services (2.13; 95% CI, 1.05 to 4.30) and for former illicit drug users versus nonusers (2.40; 95% CI, 1.05 to 5.50). CONCLUSIONS: This population-based pharmacy analysis reveals improving antiretroviral use but continuing poor pharmacy-based adherence by postpartum HIV-infected women. Receipt of HIV-focused services appears to be particularly beneficial in increasing the likelihood of treatment and adherence.

Adult↗

PubMed2000

Semiparametric regression for periodic longitudinal hormone data from multiple menstrual cycles.

We consider semiparametric regression for periodic longitudinal data. Parametric fixed effects are used to model the covariate effects and a periodic nonparametric smooth function is used to model the time effect. The within-subject correlation is modeled using subject-specific random effects and a random stochastic process with a periodic variance function. We use maximum penalized likelihood to estimate the regression coefficients and the periodic nonparametric time function, whose estimator is shown to be a periodic cubic smoothing spline. We use restricted maximum likelihood to simultaneously estimate the smoothing parameter and the variance components. We show that all model parameters can be easily obtained by fitting a linear mixed model. A common problem in the analysis of longitudinal data is to compare the time profiles of two groups, e.g., between treatment and placebo. We develop a scaled chi-squared test for the equality of two nonparametric time functions. The proposed model and the test are illustrated by analyzing hormone data collected during two consecutive menstrual cycles and their performance is evaluated through simulations.

Biometry↗

PubMed2000

A scaled linear mixed model for multiple outcomes.

We propose a scaled linear mixed model to assess the effects of exposure and other covariates on multiple continuous outcomes. The most general form of the model allows a different exposure effect for each outcome. An important special case is a model that represents the exposure effects using a common global measure that can be characterized in terms of effect sizes. Correlations among different outcomes within the same subject are accommodated using random effects. We develop two approaches to model fitting, including the maximum likelihood method and the working parameter method. A key feature of both methods is that they can be easily implemented by repeatedly calling software for fitting standard linear mixed models, e.g., SAS PROC MIXED. Compared to the maximum likelihood method, the working parameter method is easier to implement and yields fully efficient estimators of the parameters of interest. We illustrate the proposed methods by analyzing data from a study of the effects of occupational pesticide exposure on semen quality in a cohort of Chinese men.

China↗

PubMed2000

Transformation of amoxapine by Cunninghamella elegans.

We examined Cunninghamella elegans to determine its ability to transform amoxapine, a tricyclic antidepressant belonging to the dibenzoxazepine class of drugs. Approximately 57% of the exogenous amoxapine was metabolized to three metabolites that were isolated by high-performance liquid chromatography and were identified by nuclear magnetic resonance and mass spectrometry as 7-hydroxyamoxapine (48%), N-formyl-7-hydroxyamoxapine (31%), and N-formylamoxapine (21%). 7-Hydroxyamoxapine, a mammalian metabolite with biological activity, now can be produced in milligram quantities for toxicological evaluation.

Amoxapine↗

PubMed2000

LMP1 of Epstein-Barr virus induces proliferation of primary mouse embryonic fibroblasts and cooperatively transforms the cells with a p16-insensitive CDK4 oncogene.

The latent membrane protein LMP1 of Epstein-Barr virus (EBV) is often present in EBV-associated malignancies including nasopharyngeal carcinoma and Hodgkin's lymphoma. Previous work demonstrates that the LMP1 gene of EBV is sufficient to transform certain established rodent fibroblast cell lines and to induce the tumorigenicity of some human epithelial cell lines. In addition, LMP1 plays pleiotropic roles in cell growth arrest, differentiation, and apoptosis, depending on the background of the target cells. To examine the roles of LMP1 in cell proliferation and growth regulation in primary culture cells, we constructed a recombinant retrovirus containing an LMP1 gene. With this retrovirus, LMP1 was shown to stimulate the proliferation of primary mouse embryonic fibroblasts (MEF cells). It has a mitogenic activity for MEF cells, as demonstrated by an immediate induction of cell doubling time. In addition, it significantly extends the passage number of MEF cells to more than 30 after retroviral infection, compared with less than 5 for uninfected MEF cells. Furthermore, LMP1 cooperates with a p16-insensitive CDK4(R24C) oncogene in transforming MEF cells. Our results provide the first evidence of the abilities of the LMP1 gene, acting alone, to effectively induce the proliferation of primary MEF cells and of its cooperativity with another cellular oncogene in transforming primary cells.

Animals↗

PubMed2000

Biophysical injury mechanisms in electrical shock trauma.

Electrical shock trauma tends to produce a very complex pattern of injury, mainly because of the multiple modes of frequency-dependent tissue-field interactions. Historically, Joule heating was thought to be the only cause of electrical injuries to tissue by commercial-frequency electrical shocks. In the last 15 years, biomedical engineering research has improved the understanding of the underlying biophysical injury mechanisms. Besides thermal burns secondary to Joule heating, permeabilization of cell membranes and direct electroconformational denaturation of macromolecules such as proteins have also been identified as tissue-damage mechanisms. This review summarizes the physics of tissue injury caused by contact with commercial-frequency power lines, as well as exposure to lightning and radio frequency (RF), microwave, and ionizing radiation. In addition, we describe the anatomic patterns of the resultant tissue injury from these modes of electromagnetic exposures.

Animals↗

PubMed2000

DITPA prevents the blunted contraction-frequency relationship in myocytes from infarcted hearts.

Loss of the positive force-frequency relationship is a characteristic finding in failing hearts. The mechanisms of this change are not well understood. Myocardial infarction (MI) was induced in rabbits to produce left ventricular (LV) dysfunction. Beginning 1 day after MI, a subgroup of rabbits received diiodothyropropionic acid (DITPA) (3.75 mg x kg(-1) x day(-1) sc) for 3 wk. We measured contractions, Ca(2+) transients, action potentials, and sarcoplasmic reticulum (SR) Ca(2+) content at different stimulation rates in single LV myocytes. The shortening-frequency relationship was markedly flattened in MI myocytes compared with control myocytes. In addition, Ca(2+) transients, action potentials, and contractions were prolonged. Myocytes from DITPA-treated MI rabbits had preserved inotropic responses to increased stimulation rate and normal duration of action potentials and Ca(2+) transients. SR Ca(2+) content increased significantly when stimulation rate was increased from 0.5 to 2.0 Hz in control myocytes but did not change significantly in MI myocytes. Myocytes from DITPA-treated MI rabbits had a greater frequency-dependent increase in SR Ca(2+) content compared with the untreated MI rabbits. Thus single myocytes from infarcted rabbit hearts have frequency-dependent abnormalities of contractility, Ca(2+) cycling, and action potential repolarization. The flattened contraction-frequency relationship can be partially explained by an attenuation of the normal enhancement of SR Ca(2+) content that occurs when stimulation rate is increased. Chronic DITPA administration after MI largely prevents the development of these abnormalities.

Action Potentials↗

PubMed2000

Surfactant sealing of membranes permeabilized by ionizing radiation.

Acute tissue injury and subsequent inflammation, including tissue edema and erythema, can be caused by sufficiently high levels of exposure to gamma radiation. The mechanism of this tissue injury is related to the generation of reactive oxygen intermediates (ROI) which chemically alter biological molecules and cell physiology. Cell membrane lipids are vulnerable to ROI-mediated lipid peroxidation that then leads to many of the acute tissue effects. We hypothesize that increased cell membrane permeability leading to osmotic swelling and vascular transudation is one of these effects. Thus we used adult postmitotic rhabdomyocytes in culture and microscopic fluorescence techniques to quantify radiation-induced changes in cell membrane permeability. Based on time-resolved dye flux measurements, a characteristic lag time of 34 +/- 3 min was determined between exposure to 160 Gy of gamma radiation and the decrease in membrane permeability. Administration of 0.1 mM nonionic surfactant Poloxamer 188 added to the cell medium after irradiation completely inhibited the dye loss over the time course of 2 h. Thus a reproducible model was developed for studying the mechanism of acute radiation injury and the efficacy of membrane-sealing agents. As only supportive measures now exist for treating the acute, nonlethal injuries from high-dose radiation exposure, agents that can restore cell membrane function after radiation damage may offer an important tool for therapy.

Animals↗

PubMed2000

Detection of transfusion transmitted virus in hepatic and extra hepatic tissues using in situ hybridization.

OBJECTIVE: To investigate the location, distribution and significance of transfusion transmitted virus (TTV) in the liver and extra hepatic tissues. METHODS: A 123 bp fragment of TTV DNA within ORF(2) was amplified by PCR reaction and cloned into PCR2.1 vector. DNA sequencing revealed that the inserted fragment was highly isogenous with Japan ABO11494 strain. The inserted fragment was labeled with digoxigenin. Segment of liver, spleen, kidney, stomach and intestine tissues from 22 patients dead of liver diseases were detected by in situ hybridization with this digoxigenin labeled TTV DNA probe. RESULTS: TTV DNA was detected in 8 livers, 4 kidneys, 3 spleens, 2 intestines, and 2 stomachs. The positive cells of TTV DNA in livers were higher than that in other tissues. No obvious pathological topographical relationship of tissue inflammatory reaction with positive cells was found. CONCLUSION: TTV can infect hepatic and ex-hepatic tissues and may cause persistent infection.

Adolescent↗