Biomedical subjects

Z Li

Publications and source records attributed to Z Li.

At least 1,243 records · Page 69Linked to original sources

PubMed1996

Histamine-induced prolonged depolarization in rat supraoptic neurons: G-protein-mediated, Ca(2+)-independent suppression of K+ leakage conductance.

Ionic mechanisms responsible for histamine-induced prolonged depolarization in supraoptic nucleus neurons were investigated using whole-cell patch recordings in horizontally prepared brain slices from adult male rats. Bath application of histamine (1-10 microM) in control medium induced membrane depolarization in nine of 12 phasically firing, putative vasopressin cells, but not in continuous firing, putative oxytocin cells (none of five cells). Depolarization, usually accompanied by increased firing rate, started within 20 s after histamine reached the slices, lasting for 3-13 min, after which they repolarized, and this was repeatable upon washout. Chelation of intracellular Ca2+ with 11 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate and perfusion of slices with Ca(2+)-free medium blocked neither histamine-induced membrane depolarizations nor increased firing rates in 24 of 30 cells recorded. Depolarizations were always associated with decreases in membrane conductance. Following treatment with promethazine (H1 receptor antagonist) in six cells excited previously by histamine, subsequent application induced neither membrane depolarization nor increased firing. H1 receptor agonists mimicked histamine-induced depolarization (four of six cells) but the H2 receptor agonist, dimaprit (10 microM), had no effect (all of nine cells). In medium containing 0 mM Ca2+, 2 mM Co2+ and 1-2 microM tetrodotoxin, with internal Ca2+ chelation, bath application of histamine induced an apparent inward current in 15 of 20 supraoptic neurons tested. The peak of inward current evoked by 1-10 microM histamine at holding potentials around -50 mV varied from 10 to 50 pA (27.3 +/- 0.3 pA, mean +/- S.E.M.). Ramp voltage tests revealed that this inward current decreased as membrane potential was hyperpolarized and had a reversal potential of -90.1 +/- 3.8 mV (n = 10). Subtraction of current obtained before from that during histamine application revealed a current that was linear against membrane potential. Increasing external K+ concentration or introduction of K+ channel blockers in the medium attenuated or abolished histamine-induced inward current at membrane potentials close to -50 mV. When external Cl- concentration was reduced, histamine-induced inward current was still seen in five of seven supraoptic cells tested. Neither inward current nor change in conductance was observed following bath application of histamine in 11 of 12 neurons recorded using patch pipettes containing guanosine 5'-O-(2-thiodiphosphate), and in seven of eight neurons using pipettes containing guanosine 5'-O-(3-thiotriphosphate). These results suggest that histamine depolarizes supraoptic neurons, at least in part, by inhibiting a K+ leakage current mediated by H1 receptors linked to GTP-binding proteins and Ca(2+)-independent pathways. This study provides initial evidence for the second messengers regulating K+ leakage current.

Animals↗

PubMed1996

Electrophysiological properties of paraventricular magnocellular neurons in rat brain slices: modulation of IA by angiotensin II.

Whole-cell patch-clamp recordings obtained from magnocellular neurons of the hypothalamic paraventricular nucleus in brain slice preparations of adult Sprague-Dawley rats have been utilized to examine three outward potassium conductances and the ionic mechanisms through which angiotensin II exerts its neurotransmitter actions within this region. Lucifer Yellow fills showed that neurons from which we recorded had large ovoid cell bodies 11-17 microns wide and 22-35 microns long, as well as 1-3 minimally branched processes, anatomical features in accordance with those previously described for magnocellular neuroendocrine neurons. These neurons had an average resting membrane potential of -58.3 +/- 0.9 (mean +/- S.E.M.) mV, spike amplitude of 92.8 +/- 1.4 mV, and input resistance of 788.9 +/- 50.4 M omega. Most of these cells displayed irregular or continuous spontaneous activity with a mean frequency of 2.44 +/- 0.33 Hz. Voltage-clamp recordings revealed three outward potassium currents; (1) a delayed outward current (IK), (2) a Ca(2+)-dependent outward current (IK(Ca)) and (3) a transient outward current (IA). These currents were classified according to their voltage dependence, inactivation, Ca2+ dependence and pharmacology. The IK was activated by depolarization beyond -40 mV and its amplitude consistently increased with depolarizing steps. The membrane conductance underlying this current was 27.3 +/- 3.8 nS for depolarization to +50 mV. In medium containing 2 mM Ca2+, depolarization to above -20 mV evoked a slowly-activating IK(Ca) which showed minimal inactivation. This current was suppressed in Ca(2+)-free/Co2+ medium and its membrane conductance was also smaller (19.4 +/- 3.5 nS at +50 mV) than that of IK. The IA demonstrated both fast activation and inactivation and was evoked only if depolarizing pulse steps were preceded by conditioning hyperpolarization. The activation threshold was approximately -65 mV and IA amplitude increased in non-linear fashion as test voltage steps became more positive. The 90% maximum of IA conductance was 15.7 +/- 1.1 nS, and was observed at membrane potentials around -15 mV. The reversal potentials of these currents were in accordance with the K+ equilibrium potential. Tetra-ethylammonium reversibly inhibited both the peak and steady-state currents of the IK, while 4-aminopyridine suppressed the IA. Replacement of 2 mM Ca2+ with 2 mM Co2+ in our bath solution or addition of Co2+ into Ca(2+)-free medium reduced the magnitude of IA, revealing the existence of a Co(2+)-sensitive IA. Bath administration of 10(-7) M angiotensin was without significant effect on IK, but resulted in a statistically significant reduction in IA (-31.0 +/- 4.1%) in 12 of 14 paraventricular nucleus cells tested, effects which were not observed following pretreatment with the AT1 receptor antagonist losartan. We conclude that in paraventricular nucleus magnocellular cells, like other CNS neurons, at least three sets of potassium channels contribute to the outward current evoked by depolarization. Our data also demonstrate ionic mechanisms through which angiotensin may act at AT1 receptors to influence the excitability of hypothalamic neuroendocrine cells.

Angiotensin I↗

PubMed1996

Aging reduces neostriatal responsiveness to N-methyl-D-aspartate and dopamine: an in vitro electrophysiological study.

Excitatory amino acids and dopamine interact to control information flow in the neostriatum. The present study was designed to examine some of the age-induced alterations in the interaction of these two neurotransmitter systems. First, responsiveness of neostriatal neurons to glutamate and N-methyl-D-aspartate was compared in neurons from young and in aged animals. N-Methyl-D-aspartate function was chosen for emphasis because declines in cognitive processes during aging are thought to involve changes in this excitatory amino acid receptor. Second, the age-related changes in dopamine's ability to modulate responses mediated by excitatory amino acid receptors was examined. Specifically, the ability of dopamine to differentially modulate responses induced by N-methyl-D-aspartate and glutamate was assessed. There is considerable evidence for alterations in dopamine receptors and behavioral responses to dopamine in aged animals. It thus becomes important to determine how these alterations are reflected at an electrophysiological level. The responses to application of excitatory amino acid agonists and dopamine as well as changes in synaptic responses mediated by activation of N-methyl-D-aspartate receptors were assessed in 69 neurons obtained from young Fischer 344 rats (3-5 months) and young cats (3-4 years) and 69 neurons obtained from aged Fischer 344 rats (24-26 months) and aged cats (10-16 years) using an in vitro slice preparation. The results indicated that populations of aged neurons from both rats and cats displayed qualitative and quantitative alterations in responses to iontophoretic application of excitatory amino acid receptor agonists. These alterations included lack of response, unusual responses consisting of depolarizations without action potentials or combinations of prepotentials and full amplitude action potentials. Threshold currents for induction of responses were also significantly elevated in neurons from aged animals. Synaptic response components mediated by activation of N-methyl-D-aspartate receptors in aged rats were reduced as well. Exposure to Mg(2+)-free artificial cerebrospinal fluid resulted in marked increases in the size of responses evoked by local stimulation in young neurons from rats. These increases, which are mediated by activation of N-methyl-D-aspartate receptors, were significantly attenuated in aged neurons. The ability of dopamine to modulate responses mediated by activation of excitatory amino acid receptors was reduced in cells from both aged rats and cats. Subpopulations of cells were either unresponsive to dopamine or required higher iontophoretic current intensities to modulate excitatory amino acid-induced responses. The present findings further document age-induced changes in neostriatal electrophysiology indicating that interactions between excitatory amino acids and dopamine appear to be compromised during aging. They emphasize alterations in N-methyl-D-aspartate receptor function and suggest further than the ability of neostriatal neurons to integrate information is altered during aging. The present findings are supported by data from the literature indicating decreases in N-methyl-D-aspartate receptor function during aging. Furthermore, the decreases in excitatory amino acid function during aging suggest that therapeutic interventions designed to prevent or retard the deleterious effects of age in the neostriatum might be directed toward enhancing excitatory amino acid receptor function.

Age Factors↗

PubMed1996

Structure-based design of parasitic protease inhibitors.

To streamline the preclinical phase of pharmaceutical development, we have explored the utility of structural data on the molecular target and synergy between computational and medicinal chemistry. We have concentrated on parasitic infectious diseases with a particular emphasis on the development of specific noncovalent inhibitors of proteases that play a key role in the parasites' life cycles. Frequently, the structure of the enzyme target of pharmaceutical interest is not available. In this setting we have modeled the structure of the relevant enzyme by virtue of its sequence similarity with proteins of known structure. For example, we have constructed a homology-based model of falcipain, the trophozoite cysteine protease, and used the computational ligand identification algorithm DOCK to identify in compuo enzyme inhibitors including oxalic bis(2-hydroxy-1-naphthyl-methylene)hydrazide (1) [Ring, C. S.; Sun, E.; McKerow, J. H.; Lee, G.; Rosenthal, P. J., Kuntz, I. D.; Cohen, F. E., Proc. Natl Acad. Sci. U.S.A. 1993, 90, 3583]. Compound 1 inhibits falcipain (IC50 6 microM) and the organism in vitro as judged by hypoxanthine uptake (IC50 7 microM). Following this lead, to date, we have identified potent bis arylacylhydrazides (IC50 150 nM) and chalcones (IC50 200 nM) that are active against both chloroquine-sensitive and chloroquine-resistant strains of malaria. In a second example, cruzain, the crystallographically determined structure of a papain-like cysteine protease, resolved to 2.35 A, was available. Aided by DOCK, we have identified a family of bis-arylacylhydrazides that are potent inhibitors of cruzain (IC50 600 microM). These compounds represent useful leads for pharmaceutical development over strict enzyme inhibition criteria in a structure-based design program.

Aldehydes↗

PubMed1996

Presence of cell-free human immunodeficiency virus in cervicovaginal secretions is independent of viral load in the blood of human immunodeficiency virus-infected women.

OBJECTIVE: The purpose of this study was to establish virologic or molecular criteria for evaluating the rate of transmission of human immunodeficiency virus type 1 and for defining the role of virus burden in the development of gynecologic diseases in human immunodeficiency virus-infected women. STUDY DESIGN: Paired samples of blood and cervicovaginal secretions from 63 human immunodeficiency virus-seropositive women were evaluated for cell-free and cell-associated virus load by several methods, including quantitative cultures and reverse transcription polymerase chain reaction. RESULTS: All women showed evidence of virus infection in both blood and cervicovaginal secretions by a combination of in vitro culture and molecular detection methods. The CD4+ cell counts in these women ranged from < 200/microliter to > 500/microliter. Blood plasma of 26% women (12/46) did not show detectable levels of human immunodeficiency virus ribonucleic acid by reverse transcription polymerase chain reaction (< 10(2)/100 microliters). These same women had significant amounts of human immunodeficiency virus in the cell-free cervicovaginal secretions (10(2) to 10(5) copies per 100 microliters). In contrast, 17% (8/46) women with significant quantity of human immunodeficiency virus ribonucleic acid in the blood plasma had negative results for human immunodeficiency virus in the cervicovaginal secretions. Further, treatment of women with the antiviral drug zidovudine did not change the human immunodeficiency virus-1 detection rate in plasma ribonucleic acid but showed significant reduction in the ability to detect human immunodeficiency virus ribonucleic acid in cell-free cervicovaginal secretions (p = 0.036). CONCLUSIONS: We conclude that the replication kinetics of human immunodeficiency virus in the blood and cervicovaginal cells are unrelated, independent events. Further, there is no correlation between the virus load or the CD4+ cell counts in the blood and the presence or absence of quantifiable human immunodeficiency virus in cervicovaginal secretions.

Antiviral Agents↗

PubMed1996

Low salt intake down-regulates the guanylin signaling pathway in rat distal colon.

BACKGROUND & AIMS: Guanylin, an endogenous gastrointestinal peptide, causes the translocation of NaCl from interstitial fluid to the intestinal lumen. The aim of this study was to examine whether changes in dietary salt intake lead to compensatory changes in expression of the guanylin signaling pathway. METHODS: Rats received low-, normal-, or high-sodium diets for 1 week. Colonic guanylin expression was evaluated by Western and Northern blotting, rates of guanylin secretion by measuring biologically active guanylin released into the medium from colon explants, and expression of the guanylin receptor (C-type guanylate cyclase) by Northern blotting and bioassay. RESULTS: By every criterion, the low-salt diet reduced expression of guanylin to 30%-40% of the level found in control animals. Guanylin receptor expression was also decreased, although less dramatically and with a lower statistical significance. For both guanylin and guanylin receptor, the high-salt diet had no significant effect on expression. CONCLUSIONS: The data support the hypothesis that the guanylin pathway is down-regulated as an adaptive response to salt restriction.

Animals↗

PubMed1996

Impact of hydrogenated fat consumption on endogenous cholesterol synthesis and susceptibility of low-density lipoprotein to oxidation in moderately hypercholesterolemic individuals.

The effects of replacing corn oil with corn oil margarine in stick form on endogenous cholesterol synthesis and susceptibility of low-density lipoprotein (LDL) to oxidation were assessed in 14 middle-aged and elderly men and women aged 63 +/- 12 years (mean +/- SD) with moderate hypercholesterolemia (mean LDL-cholesterol [LDL-C], 4.24 +/- 0.59 mmol/L at the time of recruitment). Subjects consumed each of two diets for 32-day periods, one enriched in corn oil, which contained 30% of energy as fat (7% saturated fatty acid [SFA], 9% monounsaturated fatty acid [MUFA] [0.4% 18:1n9 trans], and 11% polyunsaturated fatty acid [PUFA]) and 85 mg cholesterol/4.2 MJ, and one enriched in stick corn oil margarine, which contained 30% fat (8% SFA, 12% MUFA [4.2% 18:1n9trans], and 8% PUFA) and 77 mg cholesterol/4.2 MJ. Both diets were isocaloric and supplied by a metabolic research kitchen. Mean total cholesterol levels were lowest (P = .039) when subjects consumed the corn oil-enriched diet (5.01 +/- 0.51 mmol/L) as compared with the margarine-enriched diet (5.30 +/- 0.58 mmol/L). LDL-C levels were 3.24 +/- 0.51 and 3.50 +/- 0.54 mmol/L when subjects consumed corn oil-and margarine-enriched diets, respectively (P = .058). There were no significant differences in high-density lipoprotein cholesterol (HDL-C) or triglyceride concentrations between the two experimental periods. Consumption of the margarine-enriched diet versus the corn oil-enriched diet tended to result in lower cholesterol fractional synthetic rates ([C-FSRs] 0.0466 +/- 0.0175 and 0.0668 +/- 0.0298, respectively, P = .080) and cholesterol absolute synthetic rates ([C-ASRs] 1.1761 +/- 0.5375 and 1.6954 +/- 0.8685, respectively, P = .092); however, differences did not reach statistical significance. Consumption of the margarine-enriched diet versus the corn oil-enriched diet resulted in a significantly higher concentration of alpha-tocopherol in both plasma and LDL(P = .004 and P = .011, respectively). LDL particle size tended to be smaller after subjects consumed the margarine-enriched diet versus the corn oil-enriched diet (P = .103). Susceptibility of LDL to oxidation was similar after consumption of the corn oil- and margarine-enriched diets. These data suggest that an increased rate of endogenous cholesterol synthesis did not contribute to the higher plasma cholesterol concentrations during the period when subjects consumed the margarine-enriched diet. Therefore, the increase in cholesterol concentration resulting from margarine consumption was likely attributable, at least in part, to a decreased catabolic rate of cholesterol. Additionally, susceptibility of LDL to in vitro oxidation was not altered by consumption of hydrogenated fat.

Adolescent↗

PubMed1996

Feasibility study of multileaf collimated electrons with a scattering foil based accelerator.

BACKGROUND AND PURPOSE: There is an ever evolving process to improve the technical aspects of electron beam delivery. Both the foil/applicator and scanning electron beam systems have gone through recent upheavals. Concomitantly, multileaf collimators are now a staple method for collimating photons. We undertook a study of multileaf collimated electron beam (MLCEB) using a dual scattering foil system. MATERIALS AND METHODS: We compared MLCEB with applicator collimated electron beams (AEB) by examining the dosimetric aspects of the two systems using 70 and 80 cm SSDs for the MLCEB, the minimum practical SSDs achievable. Percent depth dose, isodose profiles, output factors, leakage, surface dose, bremstrahlung, effective SSDs, etc. were measured using film and/or ion chamber. Clinical fields, such as posterior neck node (PNN), were compared. We also investigated the use of MLCEB for arc therapy using segments. RESULTS: In all cases, the MLCEB performed inferior, as judged by isodoses, uniformity index (UI) and penumbra analysis. The 80 cm SSD (minimum for PNN), low energy, small fields, was the worst case. For a 6 MeV beam, the UI/penumbra was 0.823/10 mm for the AEB, and 0.561/29 mm for the MLCEB at 80 cm SSD. The PNN multileaf fields exhibited narrow 90% and 80% isodose lines, and wide 20% and 10% lines. CONCLUSIONS: We conclude that multileaf for PNN fields could not be matched to adjacent off-cord photon fields. The "stair-stepping' effect associated with MLC photons was absent for electrons.

Electrons↗

PubMed1996

Inhibition of LPS-induced tumor necrosis factor-alpha production by colchicine and other microtubule disrupting drugs.

Colchicine has been shown to act as an antiinflammatory agent. In this study, we examined whether colchicine and other microtubule-depolymerizing drugs affected the production of TNF-alpha. When rat peritoneal macrophages were stimulated by LPS, addition of colchicine, vincristine, vinblastine or nocodazole was found to inhibit TNF-alpha release in a concentration-dependent manner. Suppression of TNF-alpha release was not due to interference with secretion as the cytokine did not accumulate intracellularly following colchicine treatment. Colchicine markedly enhanced PGE2 release from LPS-stimulated macrophages. However, addition of the cyclooxygenase inhibitor indomethacin only partially reversed the suppressive effect of colchicine on TNF-alpha production. Colchicine caused a strong reduction of LPS-induced TNF-alpha mRNA accumulation, suggesting that a pretranslational effect may represent the primary mechanism by which colchicine reduced TNF-alpha production. These observations could have clinical relevance in ameliorating undesirable effects due to excessive TNF-alpha production, for example following LPS stimulation of monocytes/macrophages in gram-negative sepsis. Furthermore, these drugs may provide useful tools to study the apparent involvement of the microtubular system in cytokine gene expression and cytokine production.

Animals↗

PubMed1996

Suppression of LPS-induced tumor necrosis factor-alpha gene expression by microtubule disrupting agents.

Microtubule disrupting agents such as colchicine have been shown to reduce TNF-alpha production in macrophages. To examine molecular mechanisms underlying the action of colchicine, TNF-alpha gene expression was studied in the murine macrophage cell line PU5-1.8. An LPS stimulation caused an intense up-regulation of TNF-alpha gene expression which was followed by a high TNF-alpha protein production. Simultaneous addition of colchicine (10 microM) suppressed LPS-induced TNF-alpha mRNA accumulation by one-third and TNF-alpha protein release by two-thirds. This effect was shared by vinblastine and vincristine, chemically different agents that also disrupt microtubule polymerization. For full suppressive activity on TNF-alpha gene expression, colchicine had to be present for 3 h in LPS-stimulated macrophage cultures. With nuclear run-on transcription experiments we could demonstrate that colchicine primarily inhibited de novo gene transcription and did not accelerate degradation of TNF-alpha mRNA in actinomycin D-treated macrophages. Thus, the well-known antiinflammatory action of microtubule depolymerizing agents may be largely due to a reduced TNF-alpha gene expression.

Animals↗

PubMed1996

The use of immediate frozen autogenous mandible, for benign tumour mandibular reconstruction.

Reimplantation of frozen autogenous lesioned mandible was performed in 31 patients with mandibular bone cysts and tumours. 26 reimplanted grafts survived without complications following surgery. Two cases had postoperative infections which were resolved following appropriate antibiotic treatment. Three grafts were removed due to the severe infection and wound dehiscence. The follow-up from 6 months to 4 years showed that there was no recurrence of the primary lesion in the 28 successful cases. Satisfactory facial contour after surgery was achieved in 19 patients, while other patients showed relatively mild facial disfigurement. The inferior alveolar nerve was repaired in 4 cases and a functional evaluation was made using an acupuncture pressure tester.

Adolescent↗

PubMed1996

Fatty acid unsaturation increases expression and capping of murine lymphocyte CD44 and CD45.

We studied the effect of incubating murine lymphocytes with cis-unsaturated fatty acids on expression and capping of CD44 and CD45. Lymphocytes were incubated with stearic (18:0) or oleic (18:1 omega-9) acid bound to bovine serum albumin (BSA). After incubation with rat anti-CD44 or anti-CD45 monoclonal antibodies and then with fluorescent-labeled anti-rat antibody, mean fluorescence intensity (FI) was measured by using flow cytometry. Capping was measured after warning and fixation in paraformaldehyde. Steady-state fluorescence anisotropy (rs) was measured after the cells had been incubated with trimethylammoniumdiphenylhexatriene. Incubation with oleic acid, but not stearic acid or BSA alone, was associated with an increase in FI of CD44. Expression of CD45, however, was increased by both stearic and oleic acids to the same degree over BSA controls. CD44 and CD45 capping were both increased by incubation with oleic acid. Rs was decreased in cells incubated with oleic acid, suggesting an increase in membrane fluidity. We conclude that incubation with oleic acid increases expression of CD44 and increases capping of both CD44 and CD45. These findings were confirmed in feeding experiments, in which rs was reduced and CD44 capping increased by polyunsaturated fatty acid diets.

Animals↗

PubMed1996

A conserved degradation signal regulates RAG-2 accumulation during cell division and links V(D)J recombination to the cell cycle.

The proteins RAG-1 and RAG-2 are essential for initiation of V(D)J recombination. In dividing cells, RAG-2 accumulates during G1 and is undetectable during the S and G2/M cell cycle phases. A conserved degradation signal, including an essential CDK phosphorylation site at Thr-490, regulates RAG-2 accumulation during cell division and links V(D)J recombination to the cell cycle. Mutations within this signal abolish periodic degradation of RAG-2 protein in dividing cells. In mice expressing endogenous or wild-type transgenic RAG-2, V(D)J recombination intermediates accumulate preferentially in G0/G1 thymocytes; this restriction is relieved by mutation of Thr-490 to alanine (T490A). Thus, periodic destruction of RAG-2 protein couples V(D)J recombination to cell cycle phase. Using transgenic mice expressing the T490A RAG-2 mutant and a functional T cell receptor beta chain, we demonstrate that coupling of V(D)J recombination to the cell cycle is not essential for enforcement of allelic exclusion.

Amino Acid Sequence↗

PubMed1996

Risk factors for initiation of drug use among young males in southwest China.

To identify risk factors for drug use among young males in southwest China a cross-sectional survey and a non-concurrent cohort study were conducted. Demographic, behavioral and drug-use information was collected from young males aged 18-29 years. The non-concurrent cohort included the period 1 January 1991 to 1 August 1994. A total of 1548 subjects were interviewed. The cumulative incidence increased between 1991 and 1993. Multivariate analysis identified the following significant risk factors for drug use: being divorced/widowed/separated, having been encouraged by friends/others to try drugs, smoking cigarettes, belonging to the Jingpo ethnic group, and having a family member who used drugs in 1991. More than 6 years of education was a protective factor for drug use. Drug use in the community was associated with having been encouraged by friends or others to try drugs (71%), to smoking cigarettes (50%), and to belonging to the Jingpo group (24%). The results are consistent with community based prevention approaches which should be studied carefully because of the unique cultural and epidemiological situation in China.

Adolescent↗

PubMed1996

Site-directed mutagenesis of the katG gene of Mycobacterium tuberculosis: effects on catalase-peroxidase activities and isoniazid resistance.

Recent studies examining the molecular mechanisms of isoniazid (INH) resistance in Mycobacterium tuberculosis have demonstrated that a significant percentage of drug-resistant strains are mutated in the katG gene which encodes a catalase-peroxidase, and the majority of these alterations are missense mutations which result in the substitution of a single amino acid. In previous reports, residues which may be critical for enzymatic activity and the drug-resistant phenotype have been identified by evaluating INH-resistant clinical isolates and in vitro mutants. In this study, site-directed mutagenesis techniques were utilized to alter the wild-type katG gene from M. tuberculosis at 13 of these codons. The effects of these mutations were determined using complementation assays in katG-defective, INH-resistant strains of Mycobacterium smegmatis and Mycobacterium bovis BCG. This mutational analysis revealed that point mutations in the katG gene at nine of the 13 codons can cause drug resistance, and that enzymatic activity and resistance to INH are inversely related. In addition, mutations in the mycobacterial catalase-peroxidase which reduce catalase activity also decrease peroxidase activity.

Antibodies, Bacterial↗

PubMed1996

Adrenergic regulation of ICER (inducible cyclic AMP early repressor) and beta1-adrenergic receptor gene expression in C6 glioma cells.

ICER (inducible cyclic AMP early repressor), a member of the cyclic AMP response element (CRE) modulator (CREM) family of transcription factors, is a powerful repressor of cyclic AMP-mediated transactivation. Our studies characterize the regulation of ICER in C6 glioma cells and investigate its role in repressing transcription of the beta1-adrenergic receptor (beta1AR) gene. Incubation with isoproterenol (100 nM) results in a rapid induction in levels of mRNA for ICER and its splice variant ICERgamma, with maximal induction occurring after 2 h of treatment. Incubation with isoproterenol also increased levels of CREM isoforms within 1 h; this was unexpected given previous reports that these isoforms are not rapidly induced. Increased expression of ICER and CREM was accompanied by induction of two CRE-binding complexes. The presence of ICER in these two CRE-binding complexes is demonstrated by their disruption with CREM antibody and by their comigration with recombinant ICER. Because the time course for isoproterenol induction of ICER mRNA and CRE binding corresponds to that for down-regulation of beta1AR mRNA levels in C6 glioma cells, the influence of ICER beta1AR transcription was directly examined. Coexpression of ICER significantly decreased transcriptional activity of a rat beta1AR promoter-luciferase reporter construct that contains a CRE. In contrast, coexpression of ICER did not influence two truncated rat beta1AR promoter constructs that lack the CRE site. These data demonstrate that ICER can interact at the beta1AR promoter to repress transcription.

Animals↗

PubMed1996

Health status by gastrointestinal diagnosis and abuse history.

BACKGROUND & AIMS: Standardized assessment of health status by diagnosis (functional vs. organic) and the relative influence of abuse history on health status have not been studied previously. The aim of this study was to estimate the health status and abuse history for gastrointestinal diagnoses among patients seen in a tertiary-care gastroenterology clinic and to evaluate the relative predictive effects of diagnosis and abuse history on health status. METHODS: Standardized measures of sexual and physical abuse history and six health status measures were estimated for the patients by diagnosis. Analysis of covariance was performed to determine the relative contributions of diagnosis type and abuse history on the health status measures. RESULTS: Patients with functional gastrointestinal diagnoses had poorer health status and a higher frequency of severe types of abuse than patients with structural diagnoses. Independent of abuse history, functional diagnosis was significantly associated with greater pain severity and psychological distress and poorer daily function. Independent of diagnosis, abuse history significantly contributed to greater pain severity, more days in bed, more psychological distress, and poorer daily function. CONCLUSIONS: The type of diagnosis and abuse history independently contributed to the health status of this population. Therefore, medical symptoms alone may not be sufficient to understand patients' health status. Attention must also be paid to contributing psychosocial factors.

Activities of Daily Living↗

PubMed1996

Accuracy of Monte Carlo photon transport simulation in characterizing brachytherapy dosimeter energy-response artefacts.

Practical dosimeters in brachytherapy, such as thermoluminescent dosimeters (TLD) and diodes, are usually calibrated against low-energy megavoltage beams. To measure absolute dose rate near a brachytherapy source, it is necessary to establish the energy response of the detector relative to that of the calibration energy. The purpose of this paper is to assess the accuracy of Monte Carlo photon transport (MCPT) simulation in modelling the absolute detector response as a function of detector geometry and photon energy. We have exposed two different sizes of TLD-100 (LiF chips) and p-type silicon diode detectors to calibrated 60Co, HDR source (192Ir) and superficial x-ray beams. For the Scanditronix electron-field diode, the relative detector response, defined as the measured detector readings per measured unit of air kerma, varied from 38.46 V cGy-1 (40 kVp beam) to 6.22 V cGy-1 (60Co beam). Similarly for the large and small chips the same quantity varied from 2.08-3.02 nC cGy-1 and 0.171-0.244 nC cGy-1, respectively. Monte Carlo simulation was used to calculate the absorbed dose to the active volume of the detector per unit air kerma. If the Monte Carlo simulation is accurate, then the absolute detector response, which is defined as the measured detector reading per unit dose absorbed by the active detector volume, and is calculated by Monte Carlo simulation, should be a constant. For the diode, the absolute response is 5.86 +/- 0.15 (V cGy-1). For TLDs of size 3 x 3 x 1 mm3 the absolute response is 2.47 +/- 0.07 (nC cGy-1) and for TLDs of 1 x 1 x 1 mm3 it is 0.201 +/- 0.008 (nC cGy-1). From the above results we can conclude that the absolute response function of detectors (TLDs and diodes) is directly proportional to absorbed dose by the active volume of the detector and is independent of beam quality.

Biophysical Phenomena↗