Biomedical subjects

J Bai

Publications and source records attributed to J Bai.

At least 19 recordsLinked to original sources

PubMed2001

CYP2E1-dependent toxicity and oxidative stress in HepG2 cells.

Induction of CYP2E1 by ethanol is one of the central pathways by which ethanol generates a state of oxidative stress in hepatocytes. To study the biochemical and toxicological actions of CYP2E1, our laboratory established HepG2 cell lines which constitutively overexpress CYP2E1 and characterized these cells with respect to ethanol toxicity. Addition of ethanol or an unsaturated fatty acid such as arachidonic acid or iron was toxic to the CYP2E1-expressing cells but not control cells. This toxicity was associated with elevated lipid peroxidation and could be prevented by antioxidants and inhibitors of CYP2E1. Apoptosis occurred in the CYP2E1-expressing cells exposed to ethanol, arachidonic acid, or iron. Removal of GSH caused a loss of viability in the CYP2E1-expressing cells even in the absence of added toxin or pro-oxidant. This was associated with mitochondrial damage and decreased mitochondrial membrane potential. Surprisingly, CYP2E1-expressing cells had elevated GSH levels, due to transcriptional activation of gamma glutamyl cysteine synthetase. Similarly, levels of catalase, alpha-, and microsomal glutathione transferase were also increased, suggesting that upregulation of these antioxidant genes may reflect an adaptive mechanism to remove CYP2E1-derived oxidants. While it is likely that several mechanisms contribute to alcohol-induced liver injury, the linkage between CYP2E1-dependent oxidative stress, mitochondrial injury, and GSH homeostasis may contribute to the toxic action of ethanol on the liver. HepG2 cell lines overexpressing CYP2E1 may be a valuable model to characterize the biochemical and toxicological properties of CYP2E1.

Animals↗

PubMed2001

Circulating thioredoxin suppresses lipopolysaccharide-induced neutrophil chemotaxis.

Thioredoxin (Trx), a redox enzyme with a conserved active site (Cys-32-Gly-Pro-Cys-35), is induced and secreted into circulation in response to inflammation. Studies here demonstrate that elevating Trx levels in circulation either by i.v. injection of recombinant Trx or stimulating Trx release in Trx-transgenic mice dramatically blocks lipopolysaccharide (LPS)-stimulated neutrophil migration in the murine air pouch chemotaxis model. Furthermore, we show that leukocyte recruitment induced by the murine chemokines KC/GROalpha, RANTES (regulated upon activation, normal T cell expressed and secreted), and monocyte chemoattractant protein-1 (MCP-1) is suppressed also in Trx-transgenic mice. Addressing the mechanism responsible for this suppression, we show that circulating Trx blocks (i) the LPS-stimulated in vitro activation of neutrophil p38 mitogen-activated protein kinase, (ii) the normal down-regulation of CD62L on neutrophils migrating into the LPS-stimulated air pouch, and (iii) the in vitro adhesion of LPS-activated neutrophils on endothelial cells. However, as we also show, Trx does not alter the expression of endothelial cell adhesion molecules (intercellular adhesion molecule-1, vascular cell adhesion molecule-1, CD62P, and CD62E) within 3 h. Collectively, these findings indicate that elevated levels of circulating Trx interfere with chemotaxis by acting directly on neutrophils. We discuss these findings in the context of recent studies reporting beneficial effects of acutely elevated Trx in ischemic injury and negative effects associated with chronically elevated Trx in HIV disease.

Animals↗

PubMed2001

The transmembrane protein Off-track associates with Plexins and functions downstream of Semaphorin signaling during axon guidance.

The Plexin family of transmembrane proteins appears to function as repulsive receptors for most if not all Semaphorins. Here, we use genetic and biochemical analysis in Drosophila to show that the transmembrane protein Off-track (OTK) associates with Plexin A, the receptor for Sema 1a, and that OTK is a component of the repulsive signaling response to Semaphorin ligands. In vitro, OTK associates with Plexins. In vivo, mutations in the otk gene lead to phenotypes resembling those of loss-of-function mutations of either Sema1a or PlexA. The otk gene displays strong genetic interactions with Sema1a and PlexA, suggesting that OTK and Plexin A function downstream of Sema 1a.

Alleles↗

PubMed2001

The tandem C2 domains of synaptotagmin contain redundant Ca2+ binding sites that cooperate to engage t-SNAREs and trigger exocytosis.

Real-time voltammetry measurements from cracked PC12 cells were used to analyze the role of synaptotagmin-SNARE interactions during Ca2+-triggered exocytosis. The isolated C2A domain of synaptotagmin I neither binds SNAREs nor inhibits norepinephrine secretion. In contrast, two C2 domains in tandem (either C2A-C2B or C2A-C2A) bind strongly to SNAREs, displace native synaptotagmin from SNARE complexes, and rapidly inhibit exocytosis. The tandem C2 domains of synaptotagmin cooperate via a novel mechanism in which the disruptive effects of Ca2+ ligand mutations in one C2 domain can be partially alleviated by the presence of an adjacent C2 domain. Complete disruption of Ca2+-triggered membrane and target membrane SNARE interactions required simultaneous neutralization of Ca2+ ligands in both C2 domains of the protein. We conclude that synaptotagmin-SNARE interactions regulate membrane fusion and that cooperation between synaptotagmin's C2 domains is crucial to its function.

Animals↗

PubMed2001

Tumor suppression and potentiation by manipulation of pp32 expression.

Alternative use of genes of the closely-related pp32 family is a common occurrence in human prostate cancer. pp32r1 and pp32r2, the oncogenic members of the pp32 family, are expressed in prostatic adenocarcinoma, while adjacent benign prostate continues to express pp32. This study focuses upon the role of pp32 in tumor suppression. We demonstrate that antisense inhibition of pp32 in NIH3T3 cells leads to a variety of phenotypic changes associated with transformation including reduced serum dependence and loss of contact inhibition. NIH3T3 cells with antisense-inhibited pp32 are not tumorigenic, but are markedly more susceptible to oncogenic stimuli such as ras. In contrast, constitutive expression of pp32 abolishes ras mediated transformation in vitro and tumorigenesis in vivo. These data demonstrate, from the functional aspect, that pp32 acts as a tumor suppressor. Furthermore, inactivation of pp32 function through alternative gene use may be a critical event in tumor evolution and progression.

3T3 Cells↗

PubMed2001

Multivalent anti-CCR ribozymes for stem cell-based HIV type 1 gene therapy.

HIV-1 infection of susceptible cells is mediated by the specific interaction of viral envelope glycoproteins with the cell surface CD4 receptor and a chemokine coreceptor, CCR5 or CXCR4. Individuals with a CCR5 genetic defect show resistance to HIV-1 infection, indicating that downregulation of CCR5 expression on target cells can prevent viral infection. In previous studies we demonstrated the utility of an anti-CCR5 ribozyme targeted to a single cleavage site in downregulating CCR5 expression and consequently providing resistance to viral infection. To improve on the level of downregulation we designed a construct containing an anti-CCR5 ribozyme heterotrimer (R5RbzTM) targeted to three different cleavage sites in CCR5 mRNA. In vitro tests showed that the anti-CCR5 ribozyme heterotrimer could effectively cleave the CCR5 RNA substrates to yield products of the expected sizes. This construct was introduced into various retroviral vectors for stable gene transduction. HOS.CD4/R5 cells stably transduced with this anti-CCR5 heterotrimer showed a marked reduction in the surface expression of CCR5 and a concomitant 70% reduction in macrophage-tropic viral infection. In addition, a retroviral vector containing the anti-CCR5 ribozyme heterotrimer and an anti-HIV-1 tat-rev ribozyme heterodimer was constructed. This construct also showed a similar inhibition of CCR5 surface expression and reduced infectability by the macrophage-tropic HIV-1 vector in HOS.CD4/R5 cells. The trimeric and multimeric ribozyme constructs were transduced into CD34+ hematopoietic progenitor cells to determine their effects on lineage-specific differentiation. We show that multivalent ribozyme gene-transduced hematopoietic progenitors differentiated normally into mature macrophages that bear CD14 and CD4 surface markers. Macrophages containing the transgenes expressed ribozymes, and showed resistance to M-tropic HIV-1 infection. These results provide strong support for the use of the trimeric anti-CCR5 ribozyme approach in a gene therapy setting for the treatment of HIV infection.

CCR5 Receptor Antagonists↗

PubMed2001

synaptotagmin mutants reveal essential functions for the C2B domain in Ca2+-triggered fusion and recycling of synaptic vesicles in vivo.

Synaptotagmin has been proposed to function as a Ca(2+) sensor that regulates synaptic vesicle exocytosis, whereas the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex is thought to form the core of a conserved membrane fusion machine. Little is known concerning the functional relationships between synaptotagmin and SNAREs. Here we report that synaptotagmin can facilitate SNARE complex formation in vitro and that synaptotagmin mutations disrupt SNARE complex formation in vivo. Synaptotagmin oligomers efficiently bind SNARE complexes, whereas Ca(2+) acting via synaptotagmin triggers cross-linking of SNARE complexes into dimers. Mutations in Drosophila that delete the C2B domain of synaptotagmin disrupt clathrin AP-2 binding and endocytosis. In contrast, a mutation that blocks Ca(2+)-triggered conformational changes in C2B and diminishes Ca(2+)-triggered synaptotagmin oligomerization results in a postdocking defect in neurotransmitter release and a decrease in SNARE assembly in vivo. These data suggest that Ca(2+)-driven oligomerization via the C2B domain of synaptotagmin may trigger synaptic vesicle fusion via the assembly and clustering of SNARE complexes.

Adaptor Protein Complex alpha Subunits↗

PubMed2001

Diagnosis of primary HIV-1 infection. Los Angeles County Primary HIV Infection Recruitment Network.

BACKGROUND: The optimal approach for diagnosing primary HIV-1 infection has not been defined. OBJECTIVE: To determine the usefulness of symptoms and virologic tests for diagnosing primary HIV-1 infection. DESIGN: Prospective cohort study. SETTING: A teaching hospital in Los Angeles and a university research center in San Diego, California. PATIENTS: 436 patients who had symptoms consistent with primary HIV infection. MEASUREMENTS: Clinical information and levels of HIV antibody, HIV RNA, and p24 antigen. RESULTS: Primary infection was diagnosed in 54 patients (12.4%). The sensitivity and specificity of the p24 antigen assay were 88.7% (95% CI, 77.0% to 95.7%) and 100% (CI, 99.3% to 100%), respectively. For the HIV RNA assay, sensitivity was 100% and specificity was 97.4% (CI, 94.9% to 98.9%). Fever, myalgia, rash, night sweats, and arthralgia occurred more frequently in patients with primary infection (P < 0.05). CONCLUSIONS: No sign or symptom allows targeted screening for primary infection. Although assays for HIV RNA are more sensitive than those for p24 antigen in diagnosing primary infection, they are more expensive and are more likely to yield false-positive results.

Antibodies, Viral↗

PubMed2001

Measurement of the layered compressive properties of trypsin-treated articular cartilage: an ultrasound investigation.

An ultrasound-compression system has been developed for the study of the layered biomechanical properties of articular cartilage. Cartilage specimens harvested from the bovine patella groove, with and without trypsin digestion, were tested using this system. It was noted that a large ultrasound reflection can be detected in the interface of the trypsin digestion front. This ultrasound reflection signal was used to differentiate the deformations of different portions of the cartilage throughout its depth when a load was applied. The equilibrium compression moduli of the digested, undigested and entire portions of articular cartilage were measured. The modulus of the cartilage without any digestion was 660+/-230 kPa. After 1h digestion with 1 mg ml(-1) trypsin solution, the thickness of the digested portion was 0.50+/-0.06m, and the modulus of the entire cartilage layer changed to 125+/-42kPa. The moduli of the digested and undigested portions were 58+/-24 kPa and 470+/-31 kPa, respectively. Similar results were obtained for the cartilage with trypsin digestion for 2 h.

Animals↗

PubMed2001

Computer simulation of the baroregulation in response to moderate dynamic exercise.

A baroregulation model, based on a previous pulsatile non-linear multielement cardiovascular model, is extended and used to study short-term regulation mechanisms. Using this model, the responses of several cardiovascular variables to different exercise levels are simulated and compared with the experimental data reported in the literature. The impact of physiological or pathological changes on the short-term regulation of arterial pressure under the stimulus of moderate dynamic exercise is then studied. The simulation results indicate that baroreflex feedback plays a critical role in the short-term regulation of arterial pressure. When the baroreflex gain decreases to one-third of the normal value, the response of the mean arterial pressure to moderate dynamic exercise and post-exercise recovery time increases by factors of 1.7 and 2.3, respectively. Clinical data from 36 subjects (two groups: normal and hypertensive) are collected to validate the model. Computer simulations for the hypertensive group show that the elastic modulus of the arterial vessel wall is increased by 1.5 times, and peripheral resistance is increased by 1.3 times the normal value, and the baroreflex gain decreases from 0.55 (for the normal group) to 0.40. The simulation results for normal and hypertensive groups agree well with the clinical data.

Adult↗

PubMed2001

Comparison of image reconstruction algorithms in myocardial perfusion scintigraphy.

The purpose of this study was to compare the clinical utility of two image reconstruction algorithms in myocardial perfusion SPECT (single-photon emission computed tomography): filtered back-projection (FBP) and ordered subset expectation maximization (OSEM). A rest/stress one-day protocol with 99mTc-MIBI or tetrofosmin was performed on 102 consecutive patients who underwent coronary angiography. After SPECT data acquisition, images were reconstructed with FBP and OSEM algorithms. We assessed diagnostic performance (sensitivity, specificity and accuracy) in detecting coronary artery stenosis and evaluated regional tracer uptake with a 4-point scoring system. Although there were no significant differences in diagnostic performance between FBP and OSEM reconstruction, the OSEM method yielded higher uptake in the RCA area than the FBP method by reducing the count-loss artifact due to hepatic uptake of the tracers. In addition, regional uptake in the LCX area was significantly lower in the OSEM image than in the FBP image; this phenomenon was observed mainly in patients with coronary stenosis and/or infarction in the LCX territory. In conclusion, OSEM and FBP offered comparable diagnostic performance in stress myocardial perfusion SPECT. The OSEM method contributed to reduction of the count-loss artifact in inferior and posterior walls and to easy recognition of hypoperfusion in the LCX area.

Algorithms↗

PubMed2001

Multiple modeling in the study of interaction of hemodynamics and gas exchange.

Circulation plays an important rule in gas exchange. Therefore, there is an interaction between circulation and gas exchange. To understand the dynamic effect of these two physiological systems, a computer simulation model of hemodynamics and gas exchange is established in this work. This model includes two physiological systems, namely the respiratory and circulatory systems. It consists of five parts: the model of gas transport, exchange and storage within the body, the multi-element nonlinear mathematical model of human circulatory system, an alveolar ventilation controller, a cardiac output controller, and a controller of breathing frequency. Model simulations provide results consistent with both dynamic and steady-state responses under hypoxia. Simulation results can reflect the interaction of hemodynamics and gas exchange. Using this model, the changes of pulmonary arterial pressure and right ventricular pressure in high altitude are studied. The optimal mode of breathing extra oxygen using nasal prongs or a facial mask is studied. This model may provide a useful tool to study reaction of hypoxia and the oxygen inhalation mode under hypoxia environments.

Carbon Dioxide↗

PubMed2001

Pathological changes in human malignant carcinoma treated with high-intensity focused ultrasound.

The purpose of this study was to investigate the pathologic changes of extracorporeal ablation of human malignant tumors with high-intensity focused ultrasound (HIFU). HIFU treatment was performed in the 164 patients with liver cancer, breast cancer, malignant bone tumor, soft tissue sarcoma and other malignant tumors at focal peak intensities from 5000 W x cm(-2) to 20,000 W x cm(-2), with operating frequencies of 0.8 to 3.2 MHz. To explore the pathologic impact of extracorporeal HIFU, 30 patients with malignant carcinoma underwent surgical removal after HIFU treatment. Pathologic findings showed that the treated tissues demonstrated homogeneous coagulative necrosis with an irreversible tumor cell death and severe damage to tumor blood vessels at the level of microsvasculature within the HIFU-targeted region. Thermolesions to intervening tissue were never observed. The treated region had a sharp border comprising only several cell layers between the treated and untreated areas. The repair of lesions had the processes of necrotic tissue absorption and granulation tissue replacement. It is concluded that extracorporeal treatment of human solid malignancies with HIFU could be safe, effective and feasible. As a noninvasive therapy, HIFU would be used clinically to treat patients with solid malignancies.

Bone Neoplasms↗

PubMed2001

Expression of pp32 gene family members in breast cancer.

The pp32 gene family consists of at least three closely related members, pp32, pp32r1 and pp32r2. In spite of a high degree of identity at the nucleotide level, pp32 functionally behaves as a tumor suppressor where as pp32r1 and pp32r2 are pro-oncogenic. The purpose of this pilot study was to determine pp32-related expression and whether alternative gene use among the pp32 family members occurred in human breast cancer. As a first step, in situ hybridization with a riboprobe capable of hybridizing with all the three members showed abundant pp32-related mRNA in benign ducts and acini and in infiltrating ductal carcinomas. A total of 100/102 cases were positive. Further, a detailed molecular analysis by RT-PCR, cloning, and sequencing was performed in five frozen infiltrating breast carcinomas and matched benign breast tissues. Oncogenic pp32r1 (5/5) and pp32r2 (3/5) expression was observed in carcinomas where as benign breast tissues expressed pp32. 4/5 carcinomas continued to express pp32 but one was devoid of pp32 expression. These results suggest that alternative expression of pp32 family members may be common in human breast cancer and the analysis of the profile of pp32-related expression might be helpful in understanding the role of these genes in breast cancer pathogenesis.

Breast Neoplasms↗

PubMed2001

The incidence of tuberculosis in a cohort of South-East Asian refugees arriving in Australia 1984-94.

We have used record linkage analysis to describe the incidence of tuberculosis in a cohort of 24 652 predominantly south-east Asian refugees who arrived in Sydney, Australia during the period 1984 to 1994. Cases that had been registered with the State Department of Health were confirmed by examination of case records. After an average follow-up interval of 10.3 years there were 189 cases of tuberculosis, equivalent to an average incidence rate of 74.9 cases per 100 000 person-years. The highest incidence rate was in 40-49 year olds and 47% of cases were in women. One hundred and twenty seven cases (67%) were pulmonary and, of these, 64 (50%) were direct smear positive. The incidence of tuberculosis in this cohort is similar to that observed among Vietnamese migrants to Australia and the USA and substantially higher than the incidence among people born in Australia. It is important to maintain awareness of the diagnosis of tuberculosis, especially in countries such as Australia, where the incidence in the general population is low but where there are large populations of migrants and refugees in whom a higher incidence is expected.

Adolescent↗

PubMed2001

Proliferation and differentiation of prostatic stromal cells.

OBJECTIVES: To investigate the effects of androgen, transforming growth factor beta1 (TGF-beta) and basic fibroblast growth factor (bFGF) on the proliferation and differentiation of prostatic stromal cells of the dog in vivo and human stromal cells in vitro. MATERIALS AND METHODS: Twenty-two dogs had their serum concentration of testosterone and oestradiol determined by radioimmunoassay before and after castration. Light microscopy, transmission electron microscopy and an in situ cell-death assay were carried out successively before and after castration to evaluate prostatic histomorphology. A semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to evaluate the expression of TGF-beta, bFGF and myosin in the canine prostate tissue after castration. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT), TGF-beta and bFGF in various concentrations. The proliferation of the cell cultures was detected by the tetrazolium assay. The differentiation from fibroblasts to smooth muscle cells (SMCs) was deduced by measuring the expression of SMC-specific proteins (myosin and smoothelin) using immunohistochemistry and RT-PCR. RESULTS: Castration resulted in a significant decrease in circulating testosterone levels (P < 0.01), but did not affect the circulating oestradiol levels (P > 0.05). The prostatic stromal cells, including SMCs and fibroblasts, diminished and underwent a serial pathological change of atrophy and apoptosis after castration. The atrophic cells were filled with intracellular lipofuscin. The expression of SMC myosin declined after castration, coincident with the increase in TGF-beta mRNA level and decline in bFGF mRNA. In vitro, TGF-beta inhibited the growth of human prostatic stromal cells during exponential growth, while enhancing myosin staining and stimulating the expression of smoothelin in confluent cultured stromal cells. bFGF stimulated the growth of the culture and inhibited the expression of smoothelin. DHT caused a weak increase in the proliferation and expression of SMC-specific proteins (P < 0.05). However, DHT and bFGF together stimulated the proliferation of stromal cells significantly more than either agent alone (P < 0.01). The combination of DHT and TGF-beta greatly enhanced the expression of SMC-specific proteins (P < 0.01), more strongly than either alone (P < 0.01). CONCLUSIONS: The whole prostate gland is an androgen-sensitive organ, with both the epithelium and stroma under the control of androgen. Androgen may direct the proliferation and differentiation of prostatic stromal cells by regulating the expression of TGF-beta and bFGF. Thus DHT, TGF-beta and bFGF may have important roles in regulating stromal cell homeostasis.

Animals↗

PubMed2001

Geranylgeranylacetone promotes induction and secretion of thioredoxin in gastric mucosal cells and peripheral blood lymphocytes.

Thioredoxin (TRX) is a redox-active protein which is induced by oxidative stresses and shows a variety of biological activities including cytoprotection against oxidative stress. We recently reported that geranylgeranylacetone (GGA), an anti-ulcer drug, induces TRX in rat hepatocytes. In this study, we demonstrate that GGA promotes induction and secretion of TRX in rat gastric mucosal cells and human peripheral blood lymphocytes (PBLs). Western blotting and a sensitive sandwich ELISA showed that TRX was induced by GGA in the cell lysates and culture supernatants of rat gastric mucosal RGM-1 cells and human PBLs. LDH releasing assay showed that GGA protected rat gastric mucosal RGM-1 cells from ethanol-induced cytotoxicity. Moreover, exogenous recombinant wild type TRX decreased 51Cr release from primary cultured rat gastric mucosal cells incubated with ethanol or hydrogen peroxide in a dose-dependent manner, whereas recombinant mutant TRX (C32S/C35S), in which the two cysteines were replaced with serines in its active site, did not. These results indicate that GGA promotes the induction and secretion of TRX in a variety of types of cells and suggest that induced or secreted TRX may play an important role in the cytoprotective action of GGA on gastric mucosal cells.

Animals↗

PubMed2001

Detection of circulating CEA molecules in human sera and leukopheresis of peripheral blood stem cells with E. coli expressed bispecific CEAScFv-streptavidin fusion protein-based immuno-PCR technique.

In this study, a method called bispecific CEAScFv-streptavidin fusion protein-based immuno-PCR technique will be tested experimentally. The application of the bispecific fusion protein-based immuno-PCR technique has the significant advantage that it can be readily applied in the clinical setting, as well as tested as a potential screening tool in high-risk populations of certain types of cancer.

Carcinoembryonic Antigen↗