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Biomedical subjects

G Wang

Publications and source records attributed to G Wang.

At least 631 records · Page 35Linked to original sources
PubMed1996

Calcium channels do not play a role in the steroid response to ACTH IN Y1 adrenocortical cells.

Y1 cells derived from mouse tumor zona fasciculata cells (ZF) were used to assess the importance of extracellular (EC) calcium availability via voltage-dependent calcium channels (VDCC) on steroidogenesis. The steroidogenic response to ACTH was investigated in the presence of blockers known to affect both calcium and potassium channels in Y1 cells. Y1 cells respond to either ACTH (100 pM) or cAMP (300 microM) at low EC Ca2+ (1 microM) suggesting that EC Ca2+ is not absolutely necessary for a steroidogenic response. However, increases in Ca2+ from 0.05-2.2 mM induced a small but significant biphasic response, first stimulating then inhibiting steroidogenesis. Nickel and amiloride, blockers of T-type Ca2+ channels in Y1 cells, did not depress ACTH-induced steroidogenesis. The dihydropyridine, nifedipine, which is an L-type channel antagonist did not affect ACTH-induced steroidogenesis while the agonist, Bay K 8644 was consistently inhibitory. Neither pimozide nor omega-conotoxin which suppressed Ca2+ currents in Y1 cells inhibited ACTH-induced steroidogenesis. Depolarization of the membrane which would activate VDCC's was inhibitory rather than stimulatory. The present studies using blockers of both voltage-dependent Ca2+ and K+ channels suggest that EC Ca2+ plays a modulatory role in ACTH-induced steroidogenesis in Y1 cells but the data do not support the concept that activation of voltage dependent calcium channels are an important mechanism for steroidogenesis.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1996

Concomitant low dose-rate irradiation and cisplatin treatment in ovarian carcinoma cell lines sensitive and resistant to cisplatin treatment.

Human ovarian carcinoma parental and cisplatin-resistant cells were evaluated for their radiation sensitivity to high and low dose-rate irradiation and for the effectiveness of cisplatin in radiosensitization. The cisplatin resistant variant A2780cp showed increased radiation resistance for both low dose-rate (LDRI) and high dose-rate irradiation. For cisplatin treatment for 1 h before and after HDRI there was radiosensitization in only the cisplatin-resistant variant. Concomitant cisplatin treatment during LDRI resulted in radiosensitization in both cell lines with dose-modifying factors ranging from 1.6 to 5.8. In this case greater radiosensitization was achieved in the parental cell line. In both cell lines the dose-modifying factors were larger when the cisplatin was refreshed every 6 h instead of 12 h during LDRI. These data show that cisplatin may be a very effective radiosensitizer when given during LDRI which is used in brachytherapy.

Antineoplastic Agents↗
PubMed1996

Murine cells transfected with human Hsp27 cDNA resist TNF-induced cytotoxicity.

Hyperthermia sensitizes tumor cells to killing by tumor necrosis factor-alpha (TNF). Sensitization is greater in cells exposed to TNF before heating begins than with the reverse sequence, and heat-shock proteins (hsp) have been suggested to protect cells from TNF cytotoxicity. Here we examined the role of Hsp27 in TNF resistance. Murine L929 cells were stably transfected with the vector pRc/CMV constitutively to express an inserted human hsp27 complementary DNA (cDNA) sequence. Parental cells produced no detectable murine homolog to human hsp27. Hsp27-sense clones expressed hsp27 messenger RNA (mRNA) and protein at 37 degrees C. Cells transfected with the cDNA in the anti-sense orientation produced anti-sense mRNA but no protein, and cells transfected with the vector alone produced neither product. Expression of hsp27 conferred significant resistance to TNF cytotoxicity in both neutral red cytotoxicity and clonogenic survival assays. Vector along and hsp27 anti-sense transfectants had a TNF response similar to that of parental L929 cells. Kinetic studies in L929 cells showed that hsp27-expressing clones exhibited resistance relative to parental cells beginning 6 h after TNF exposure, and this differential response increased by 12 and 24 h. Addition of actinomycin D to the TNF cytotoxicity assays accelerated the cytotoxicity development in parental and transfected cells, but the hsp27-sense clones were still more resistant. Hsp27-sense clones of L929 cells were also resistant to oxidative stress induced by menadione and released less arachidonic acid in response to TNF induction. These results show that hsp27 can negatively regulate the TNF cytotoxic mechanism.

Animals↗
PubMed1996

A low molecular weight outer-membrane protein of Escherichia coli O157:H7 associated with adherence to INT407 cells and chicken caeca.

Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 and O26:H11 have been shown to produce a low mol.wt outer-membrane protein (OMP) that is unique to a few serotypes of E. coli. A mutant (A10) of E. coli O157:H7 strain HA1 deficient in the OMP was constructed by TnphoA mutagenesis and assayed for its adherent ability. Adherence of A10 to intestinal epithelial cells (INT407) was significantly less than that of its parent strain (HA1). Adherence of HA1 to INT407 cells was significantly decreased by treatment with a monoclonal antibody (4E8C12) that specifically binds to the OMP. When chickens were infected experimentally with E. coli O157:H7 strains, the average number of cfu of strain A10 recovered from chicken caeca was significantly less than those of strain HA1 and wild-type strain 932 at 14 and 21 days after peroral inoculation. These data suggest that the OMP of EHEC is associated with adherence of E. coli O157:H7 to epithelial cells in vitro and chicken caeca in vivo.

Animals↗
PubMed1996

Unwrapping Cochlear implants by spiral CT.

Multielectrode, intracochlear implants were designed for individuals with profound sensorineural hearing loss who derive little or no benefit form acoustic hearing aids. Determination of each electrode's position in a patient's inner ear may improve speech processor programming to maximize speech recognition. In this paper, an approach is described to use as input a volumetric spiral computed tomography (CT) image of the Nucleus electrode array (Cochlear Pty. Ltd, Lane Cove, NSW, Australia) to unwrap it, and to measure its implanted length given starting and end points. Representative curvilinear structures were digitally synthesized in image volumes of isotropic 0.1-mm voxels. The electrode array was spirally CT-scanned in vitro and in vivo, and reconstructed on an isotropic grid in 0.1-mm steps. Two algorithms were constructed to track and measure these curvilinear structures. The first algorithm is Karhunen-Loeve (K-L)-transform based, in which the K-L transform is locally applied at a current main axis position to determine the eigenvectors of the main axis voxels, the next main axis position is estimated from the current position along the principal eigendirection, adjusted to the mass center of the orthogonal cross section passing through the estimated position, and then scaled to have a prespecified step. The second algorithm is similar to the first one but avoids use of the K-L transform. In the second algorithm, the next position is directly estimated along the local direction and then processed with the same correction and scaling operations. With user-specified starting and end points as well as a local direction at the starting point, a curvilinear structure can be automatically tracked using either of the algorithms. The first algorithm is more robust, while the second one is more efficient. In the numerical and in vitro studies, the lengths of the curvilinear structures were accurately measured. Given local directions determined in the tracking process, an electrode array image can be unwrapped into a linear array with the central electrode axis as the abscissa. The unwrapping approach allows longitudinally and cross-sectionally accurate measurement and better visualization of cochlear implant images. With preimplantation knowledge of length, width, and center electrode distance, the position of individual electrodes can be estimated after unwrapping.

Algorithms↗
PubMed1996

Conformational studies of an amphipathic peptide corresponding to human apolipoprotein A-II residues 18-30 with a C-terminal lipid binding motif EWLNS.

A peptide was designed and synthesized to enhance the lipid binding properties of a 13-residue fragment of apolipoprotein A-II. The peptide, VTDYGKDLMEKVKEWLNS [apoA-II(18-30)+], contains a five-residue amphipathic motif, EWLNS, at the C-terminus of apolipoprotein A-II residues 18-30. The lipid binding properties of apoA-II(18-30)+ were assessed using optical spectroscopy in the presence of sodium dodecyl sulfate (SDS), dodecylphosphocholine (DPC), tetradecyltrimethyl ammonium chloride (TMA) and dimyristoylphosphatidylcholine (DMPC). The fluorescence emission spectra and the circular dichroism data suggested that apoA-II(18-30)+ interacted most strongly with SDS and most weakly with DMPC. An ensemble of structures for apoA-II(18-30)+ in aqueous solution containing SDS was calculated using distance geometry/simulated annealing methods from 308 NOE-based distance restraints. The backbone (N-C-C = O) RMSD from the average structure of an ensemble of 15 out of 20 calculated structures was 0.54 +/- 0.16 A. Apart from some dynamic fraying at both termini, the distance geometry and simulated annealing calculations showed that apoA-II(18-30)+ adopted a well defined amphipathic helix with distinct hydrophobic and hydrophilic faces.

Amino Acid Sequence↗
PubMed1996

Fate of enterohemorrhagic Escherichia coli O157:H7 in bovine feces.

Dairy cattle have been identified as a principal reservoir of Escherichia coli O157:H7. The fate of this pathogen in bovine feces at 5, 22, and 37 degrees C was determined. Two levels of inocula (10(3) and 10(5) CFU/g) of a mixture of five nalidixic acid-resistant E. coli O157:H7 strains were used. E. coli O157:H7 survived at 37 degrees C for 42 and 49 days with low and high inocula, respectively, and at 22 degrees C for 49 and 56 days with low and high inocula, respectively. Fecal samples at both temperatures had low moisture contents (about 10%) and water activities ( < 0.5) near the end of the study. E. coli O157:H7 at 5 degrees C survived for 63 to 70 days, with the moisture content (74%) of feces remaining high through the study. Chromosomal DNA fingerprinting of E. coli O157:H7 isolates surviving near the completion of the study revealed that the human isolate strain 932 was the only surviving strain at 22 or 37 degrees C. All five strains were isolated near the end of incubation from feces held at 5 degrees C. Isolates at each temperature were still capable of producing both verotoxin 1 and verotoxin 2. Results indicate that E. coli O157:H7 can survive in feces for a long period of time and retain its ability to produce verotoxins. Hence, bovine feces are a potential vehicle for transmitting E. coli O157:H7 to cattle, food, and the environment. Appropriate handling of bovine feces is important to control the spread of this pathogen.

Animals↗
PubMed1996

Role of mismatch repair in the Escherichia coli UVM response.

Mutagenesis at 3,N4-ethenocytosine (epsilonC), a nonpairing mutagenic lesion, is significantly enhanced in Escherichia coli cells pretreated with UV, alkylating agents, or H2O2. This effect, termed UVM (for UV modulation of mutagenesis), is distinct from known DNA damage-inducible responses, such as the SOS response, the adaptive response to alkylating agents, or the oxyR-mediated response to oxidative agents. Here, we have addressed the hypothesis that UVM results from transient depletion of a mismatch repair activity that normally acts to reduce mutagenesis. To test whether the loss of mismatch repair activities results in the predicted constitutive UVM phenotype, E. coli cells defective for methyl-directed mismatch repair, for very-short-patch repair, or for the N-glycosylase activities MutY and MutM were treated with the UVM-inducing agent 1-methyl-3-nitro-1-nitrosoguanidine, with subsequent transfection of M13 viral single-stranded DNA bearing a site-specific epsilonC lesion. Survival of the M13 DNA was measured as transfection efficiency, and mutation fixation at the lesion was characterized by multiplex sequencing technology. The results showed normal UVM induction patterns in all the repair-defective strains tested. In addition, normal UVM induction was observed in cells overexpressing MutH, MutL, or MutS. All strains displayed UVM reactivation, the term used to describe the increased survival of epsilonC-containing DNA in UVM-induced cells. Taken together, these results indicate that the UVM response is independent of known mismatch repair systems in E. coli and may thus represent a previously unrecognized misrepair or misreplication pathway.

Cytosine↗
PubMed1996

Characterization of cytotoxic, hemolytic Aeromonas caviae clinical isolates and their identification by determining presence of a unique hemolysin gene.

Aeromonas caviae has recently been recognized as an important enteropathogen and its hemolysin is purported to be one of the virulence factors, In this study, a total of 80 clinical isolates of Aeromonas spp. were investigated by PCR with synthetic oligonucleotides targeting a cloned hemolysin-encoding sequence from an A. caviae isolate of clinical origin. Of the 35 clinical A. caviae isolates tested, only 6 contained the target sequence.

Aeromonas↗
PubMed1996

In vivo sequence diversity of the protease of human immunodeficiency virus type 1: presence of protease inhibitor-resistant variants in untreated subjects.

We have evaluated the sequence diversity of the protease human immunodeficiency virus type 1 in vivo. Our analysis of 246 protease coding domain sequences obtained from 12 subjects indicates that amino acid substitutions predicted to give rise to protease inhibitor resistance may be present in patients who have not received protease inhibitors. In addition, we demonstrated that amino acid residues directly involved in enzyme-substrate interactions may be varied in infected individuals. Several of these substitutions occurred in combination either more or less frequently than would be expected if their appearance was independent, suggesting that one substitution may compensate for the effects of another. Taken together, our analysis indicates that the human immunodeficiency virus type 1 protease has flexibility sufficient to vary critical subsites in vivo, thereby retaining enzyme function and viral pathogenicity.

Amino Acid Sequence↗
PubMed1996

Prolactin levels and antinuclear antibody profiles in women tested for connective tissue disease.

Hyperprolactinemia has been reported in some patients with active systemic lupus erythematosus (SLE). To determine if there was an association between selected autoantibodies and hyperprolactinemia, we assayed prolactin concentrations in sera from women submitted to a reference antinuclear antibody laboratory. Autoantibody-positive samples were separated into groups that contained antibodies to double-stranded DNA (anti-DNA), antibodies to SSA/Ro (anti-SSA/Ro), or antibodies to both SSA/Ro and SSB/La (anti-SSA/Ro-SSB/La). Results were compared with autoantibody-negative sera from age-matched women, submitted to the same laboratory. We also compared the study groups with a separate cohort of 84 healthy women who were not referred for autoantibody testing. Elevated prolactin levels were clustered in 20% of sera from anti-DNA-positive women < or = 50 years of age. Twenty-one percent of anti-SSA/Ro-SSB/La-positive women < 50 years of age were hyperprolactinemic. Four of the 15 hyperprolactinemic women identified in this survey had no known cause of elevated prolactin. In the other 11 individuals secondary causes such as hypothyroidism, pregnancy, chronic renal failure, and medications may have accounted for high serum prolactin values. We also examined sera by Western blot, to determine if immunoblot patterns were associated with elevated serum prolactin concentrations. The hyperprolactinemic sera yielded novel bands migrating at 70 kd, 32 kd, and 16.5 kd. This study confirmed the reported associations of hyperprolactinemia with SLE and Sjögren's syndrome. Multiple factors appeared to contribute to elevated serum prolactin levels in women with connective tissue diseases, and the presence of hyperprolactinemia was related to unique findings on immunoblot analysis.

Adolescent↗
PubMed1996

Immunohistochemical localization of mouse Clara cell 10-KD protein using antibodies raised against the recombinant protein.

To investigate the developmental regulation of the mouse Clara cell 10-KD protein (mCC10), we raised an antibody against the recombinant mCC10 protein. The coding region for the mature mCC10 protein was placed in frame with the glutathione-S-transferase gene in the pGEX2-T bacterial expression vector. The GST-mCC10 fusion protein was expressed in E. coli DH5 alpha cells. The fusion protein was purified and eluted using glutathione-Sepharose beads. The GST-mCC10 fusion protein was injected into rabbits to raise antibodies. The rabbit anti-mCC10 antibody was tested by immunoblot analysis using both purified protein as well as extracts of lung, liver, and uterus. The antibodies produced were used in immunohistochemistry and immunoelectron microscopy to detect the cellular localization of this protein in the above organs. This anti-mCC10 antibody will be useful for future investigation of the developmental biology of the lung.

Animals↗
PubMed1996

Glucocorticoid stimulation of pro-ANF mRNA expression of adult rat hypothalamic neurons in culture: a cAMP-dependent effect.

We describe here long term primary cultures of adult rat hypothalamic cells, a novel system which we have exploited to examine the effect of glucocorticoids on adult ANF neurons in vitro. ANF neurons with and without neurite outgrowths were identified by immunocytochemistry and by in situ hybridization studies, in which pro-ANF mRNA signals were localised in cultured cells. Dexamethasone (DM) augmented the abundance of pro-ANF mRNA signals in a dose-related manner with ED50 of 5 x 10(-12)M and Emax of 10(-10)M. This stimulation effect was mimicked by corticosterone but not by deoxycorticosterone. Similarly, activation of the adenylyl cyclase-cAMP system with forskolin, 8-brom-cAMP or 3-isobutyl-1-methylxanthine (IBMX), also markedly elevated the level of pro-ANF mRNA abundance. Interestingly, whereas forskolin-induced stimulation was reversed by Rp-cAMP, a cAMP antagonist, the augmentation induced by DM alone or by DM and forskolin together, was similarly suppressed by the chemical. We therefore conclude that (i) adult ANF neurons are capable of regenerating neurite outgrowths in vitro, (ii) cAMP dependent pathways play a fundamental role in regulating the expression of pro-ANF mRNA, and (iii) the stimulatory effect of glucocorticoids is indirect and mediated, at least in part, through modulating the activity of cAMP dependent pathways.

Animals↗
PubMed1996

Ghosting of pulmonary nodules with respiratory motion: comparison of helical and conventional CT using an in vitro pediatric model.

OBJECTIVE: The study was designed to compare helical CT with varying pitch and reconstruction intervals and conventional CT for revealing pulmonary nodules in a model that simulates respiratory motion in children. MATERIALS AND METHODS: CT scans were obtained in an experimental model with one nodule (3 or 10 mm) in each scan. One-second scans were obtained at rates of 10, 20, and 30 respirations per minute using conventional CT with 4-mm collimation and table incrementation and helical CT with 4-mm collimation and either 4-mm/sec (pitch, 1:1) or 8-mm/sec (pitch, 2:1) table speed. Reconstructions were at 1-, 2-, and 4-mm intervals for scans obtained using 4-mm/sec table speed and at 1- and 4-mm intervals for scans obtained using 8-mm/sec table speed. Images were independently reviewed by three radiologists who estimated the number of nodules on each image. RESULTS: Ghosting (depiction of more than one nodule in a study) was seen in 79%, 80%, and 75% of helical CT scans obtained with a 1:1 pitch using 1-, 2-, and 4-mm reconstruction intervals, respectively. By comparison, ghosting was seen in only 54% and 58% of helical CT scans with a 2:1 pitch using 1-mm reconstruction intervals and 4-mm reconstruction intervals, respectively, and in 56% of conventional CT scans (p < .0001). A single nodule was detected on all other scans, and at least one nodule was seen on all scans. CONCLUSION: Ghosting of nodules is common in this model. Ghosting was seen less often on conventional scans and helical scans with 2:1 pitch than it was on helical scans with 1:1 pitch. Nonetheless, ghosting was seen on more than 50% of all scans with each technique.

Artifacts↗
PubMed1996

A simple model for predicting the response of chicks to dietary enzyme supplementation.

Two experiments with Leghorn chicks and data from five publications were analyzed to determine whether a simple general equation could be used to predict the relationship between the amount of a crude enzyme added to a diet and chick performance. The maximum improvements in weight gain and feed: gain ratio in Leghorn chicks fed rye diets containing different concentrations of enzymes were as high as 61 and 42%, respectively. Regression analyses demonstrated that there was usually a high linear correlation (r2 > 0.9, P < 0.05) between the concentration of the enzyme when transformed into its logarithmic value and weight gain or the feed:gain ratio. The general prediction equation was Y = A + B(logX), where Y is the performance value (i.e., weight gain, grams), A is the intercept (y-axis), B is the slope of the line (change in performance per units of enzyme in the diet), and X is the amount of enzyme in the diet. The slope of the line provides an index of the overall efficacy of the enzyme treatment. The log-linear model shows that for every ninefold increase in amount of enzyme in the diet (i.e., when the amount was increased to 10 times the starting amount), there was only a doubling of improvement in chick performance. High correlations (r2 values) were also obtained when data from the literature were analyzed. The equation was applicable to different classes and ages of poultry fed diets containing rye, wheat, barley, and lupins. These studies demonstrate that there is a linear relationship between the amount of enzyme added to the diet, when expressed as a logarithmic value, and the corresponding performance of chickens.

Animal Feed↗