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Biomedical subjects

A Ray

Publications and source records attributed to A Ray.

At least 73 records · Page 4Linked to original sources
PubMed1997

Transcription factor GATA-3 is differentially expressed in murine Th1 and Th2 cells and controls Th2-specific expression of the interleukin-5 gene.

Interleukin-5 (IL-5), which is produced by CD4(+) T helper 2 (Th2) cells, but not by Th1 cells, plays a key role in the development of eosinophilia in asthma. Despite increasing evidence that the outcome of many diseases is determined by the ratio of the two subsets of CD4(+) T helper cells, Th1 and Th2, the molecular basis for Th1- and Th2-specific gene expression remains to be elucidated. We previously established a critical role for the transcription factor GATA-3 in IL-5 promoter activation in EL-4 cells, which express both Th1- and Th2-type cytokines. Our studies reported here demonstrate that GATA-3 is critical for expression of the IL-5 gene in bona fide Th2 cells. Whereas mutations in the GATA-3 site abolished antigen- or cAMP-stimulated IL-5 promoter activation in Th2 cells, ectopic expression of GATA-3 in Th1 cells or in a non-lymphoid, non-IL-5-producing cell line activated the IL-5 promoter. During the differentiation of naive CD4(+) T cells isolated from T cell receptor transgenic mice, GATA-3 gene expression was up-regulated in developing Th2 cells, but was down-regulated in Th1 cells, and antigen- or cAMP-activated Th2 cells (but not Th1 cells) expressed the GATA-3 protein. Thus, GATA-3 may play an important role in the balance between Th1 and Th2 subsets in immune responses. Inhibition of GATA-3 activity has therapeutic potential in the treatment of asthma and other hypereosinophilic diseases.

Animals↗
PubMed1997

Selective up-regulation of cytokine-induced RANTES gene expression in lung epithelial cells by overexpression of IkappaBR.

We previously reported the cloning of a cDNA for IkappaBR (for IkappaB-related) from human lung alveolar epithelial cells. IkappaBR is related to the IkappaB proteins that function as regulators of the NF-kappaB family of transcription factors. Here, we investigated the consequence of IkappaBR overexpression on the expression of NF-kappaB-regulated chemokine genes in lung alveolar epithelial cells. Chemokines play an important role in many inflammatory diseases such as asthma and AIDS. Overexpression of IkappaBR in the lung cells resulted in a rapid 50-100-fold greater production of the RANTES (regulated upon activation, normal T expressed and presumably secreted) protein upon cytokine-induction compared with control cells. IkappaBR overexpression, however, did not enhance interleukin-8 or MIP-1alpha gene expression, despite the fact that the expression of all three chemokine genes are regulated by NF-kappaB. The up-regulation of RANTES gene expression resulting from overexpression of IkappaBR correlated with increased amounts of a unique RANTES-kappaB binding activity and decreased binding of p50 homodimers. Previous studies have shown that p50 homodimers function as repressors of certain kappaB sites. Our studies suggest that IkappaBR can aid activation of select NF-kappaB-regulated genes by sequestering p50 homodimers.

Base Sequence↗
PubMed1997

Repression of interleukin-6 gene expression by 17 beta-estradiol: inhibition of the DNA-binding activity of the transcription factors NF-IL6 and NF-kappa B by the estrogen receptor.

The cytokine interleukin-6 (IL-6), a key mediator of immune and acute phase responses of the liver, has also been implicated in uterine functions. Estrogens are potent repressors of IL-6 production by uterine stromal cells. In the endometrial adenocarcinoma cell line Ishikawa, phorbol ester-induced activation of the IL-6 promoter was inhibited to basal levels by 17 beta-estradiol (E2) in a wild-type receptor-dependent fashion. Although tamoxifen has been shown to have estrogenic effects on the endometrium, it did not inhibit induction of the IL-6 promoter. We previously showed that inhibition of IL-6 gene expression by E2 does not involve high-affinity binding of the estrogen receptor (ER) to IL-6 DNA. We now report that the ER can directly interact with the transcription factors NF-IL6 and NF-kappa B and can inhibit their ability to bind DNA which might be the molecular basis for repression of IL-6 gene expression by estrogens.

Adenocarcinoma↗
PubMed1997

Involvement of an SAF-like transcription factor in the activation of serum amyloid A gene in monocyte/macrophage cells by lipopolysaccharide.

Serum amyloid A (SAA) has been linked to atherosclerosis because of its ability to remodel high-density lipoprotein by the depletion of apolipoprotein A1, its ability to bind cholesterol, and its presence in the atherosclerotic plaques of coronary and carotid arteries. In the present study, we investigated the induction mechanism of SAA gene in THP-1 monocyte/macrophage cells which play a critical role in the development of atherosclerotic fatty streak and plaque formation. We and others have shown that SAA gene is induced in monocyte/macrophage cells by lipopolysaccharide (LPS). By promoter function analysis, we show that the SAA promoter sequence between -280 and -226 can confer LPS responsiveness. Gel electrophoretic mobility shift assay detected an induced DNA-binding activity in these cells in response to LPS. Characterization of the DNA-binding protein by UV cross-linking, Southwestern blot, and antibody ablation/supershift assays revealed that it is similar to a recently reported nuclear factor designated SAF. These results demonstrated that LPS-mediated SAA gene induction in monocyte/macrophage cells is primarily due to the induction of SAF activity.

Amyloid↗
PubMed1997

Influence of stress on DDT-induced humoral immune responsiveness in mice.

The effects of different durations/intensities of stress on DDT-induced modulation of humoral immune response were evaluated in mice. DDT (20, 50, or 100 ppm x 4 weeks) per se did not influence the primary antibody response to sheep red blood cells (SRBC). However, when DDT-pretreated mice were exposed to single and multiple sessions of restraint stress (RS), the anti-SRBC antibody titers were lower than the control values, the most prominent effects being seen after 50 and 100 ppm DDT exposure in combination with a single intense stressor (24 hr RS) or repeated stress (1 hr RS x 5). In the splenic plaque forming cells (PFC) assay, similar potentiations of DDT-induced immune suppression were seen at 50 and 100 ppm exposure levels in combination with 24 hr RS or 1 hr RS (x5) procedures. In addition, the 20 ppm DDT exposure effect was also potentiated in combination with the multiple RS model. Further, other forms of stress viz. 3 hr cold restraint stress (CRS) or 6 hr RS, which per se did not influence the antibody titer or PFC response, suppressed humoral immune responses, when combined with 100 ppm DDT exposure. These results are discussed in light of the possible interactions between physical/emotional and environmental/xenobiotic stressors in the regulation of humoral immune response.

Animals↗
PubMed1997

Effects of in-vivo generation of oxygen free radicals on immune responsiveness in rabbits.

Oxygen free radicals (OFRs) generated during biological processes are reportedly involved in the pathogenesis of several disease states and various reports have indicated that oxidative stress may alter immune competence. Hence, effects of in-vivo generation of OFRs by using xanthine/xanthine oxidase (X/XO) system on immune responsiveness were evaluated in rabbits. Intravenous injections of xanthine (0.14 mg/kg) along with xanthine oxidase (2 U/Kg) following primary and secondary immunizations of animals with sheep red blood cells (SRBC) significantly attenuated the primary and secondary antibody responses respectively. In tests for cell-mediated immunity, tuberculin sensitivity and leucocyte migration inhibition were also decreased significantly in sensitized animals following X/XO treatment. The observed changes in both humoral and cell-mediated immune responses following such in-vivo generation of OFRs indicate a possible nexus between OFR generation and immune suppression.

Animals↗
PubMed1997

Anabolic androgenic steroids increase beta-endorphin levels in the ventral tegmental area in the male rat brain.

The levels of beta-endorphin and Met-enkephalin-Arg-Phe (MEAP) immunoreactivity in various brain regions (including amygdala, cortex, hippocampus, hypothalmus, nucleus accumbens, pituitary and ventral tegmental area) were studied in male rats subjected to daily intramuscular injections during 14 days of high doses (5 and 15 mg/kg) of the anabolic androgenic steroid (AAS), nandrolone decanoate. At the nandrolone dose of 15 mg/kg a significant (about 20-fold) increase in beta-endorphin levels in the ventral tegmental area (VTA) was observed. The steroid did not significantly affect the concentration of the peptide at any dose in other brain areas examined. The levels of MEAP remained unaltered in all studied regions. A slight increase in serum concentrations of both peptides was also found but this elevation was not statistically significant. The observed increase in beta-endorphin in VTA was suggested to be involved in a mechanism by which the steroid may influence the reward system in the brain. An opioid mediated stimulation of the reward system following injection of AAS supports a previous hypothesis that AAS may induce psychological dependence.

Anabolic Agents↗
PubMed1997

Serum amyloid A gene expression level in liver in response to different inflammatory agents is dependent upon the nature of activated transcription factors.

Serum amyloid A (SAA) is highly induced during many inflammatory episodes. The induction mechanism in response to turpentine and lipopolysaccharide (LPS), two major inducers of this gene, was investigated. Here we present evidence that although both agents triggered expression, SAA mRNA synthesized in the turpentine-injected rabbit liver is many-fold higher compared to that found in LPS-injected rabbit liver. We demonstrate that differential level of activation of C/EBP and NF-kappaB that interact with the proximal promoter of SAA gene is responsible for the differential expression. A very high level of C/EBP induction with little or no activation of NF-kappaB factors was noted when turpentine was used as the inducer. LPS, on the other hand, activated NF-kappaB and C/EBP, which were detected only at the early phase of induction process. These results indicate that different pathways might be activated for the regulation of hepatic expression of SAA by different inflammatory agents. One of the pathways, triggered by LPS, requires participation of both NF-kappaB and C/EBP. A second pathway, triggered by turpentine, involves only C/EBP family of transcription factors.

Animals↗
PubMed1997

Pollen tube guidance by the female gametophyte.

In flowering plants, pollen grains germinate on the pistil and send pollen tubes down the transmitting tract toward ovules. Previous genetic studies suggested that the ovule is responsible for long-range pollen tube guidance during the last phase of a pollen tube's journey to the female gametes. It was not possible, however, to unambiguously identify the signaling cells within an ovule: the haploid female gametophyte or the diploid sporophytic cells. In an effort to distinguish genetically between these two possibilities, we have used a reciprocal chromosomal translocation to generate flowers wherein approximately half the ovules do not contain a functional female gametophyte but all ovules contain genotypically normal sporophytic cells. In these flowers, pollen tubes are guided to the normal but not to the abnormal female gametophytes. These results strongly suggest that the female gametophyte is responsible for pollen tube guidance, but leave open the possibility that the gametophyte may accomplish this indirectly through its influence on some sporophytic cells.

Arabidopsis↗
PubMed1997

Effects of stress on gamma glutamyl transpeptidase (GGT) activity in lymphoid system of rats: modulation by drugs.

Effects of stress and its modulation by adaptogens were evaluated on gamma glutamyl transpeptidase (GGT) activity in different tissues of the lymphoid system in rats. Restrain stress (RSx5) suppressed the GGT activity in different tissues of lymphoid system viz. the lymphocyte, the spleen, the thymus and the macrophage, and the maximum effect was seen in the spleen. Chlordiazepoxide, a prototype anti-stress agent, which did not alter GGT activity per se, reversed the effect of RS on this enzyme activity in tissues of lymphoid system studied. Azardirachta indica (Al, Neem), an indigenous adaptogen stimulated the GGT activity per se and nearly normalised RS induced suppression of GGT in lymphoid system. The observed suppression of GGT activity in lymphoid system by stress and its modulation by natural and synthetic adaptogens indicates that GGT could be a consistent cellular/biochemical marker of stress responsiveness and stress-induced immunomodulation.

Animals↗
PubMed1997

Modulation of gamma glutamyl transpeptidase activity in lymphoid system by organochlorine pesticides in rats.

Effects of 1,1,1,-trichloro-2,2-bis (p-chlorophenyl) ethane (DDT) and lindane were studied on gamma glutamyl transpeptidase (GGT) activity in different tissues of the lymphoid system in rats. DDT (100 or 200 ppm) and lindane (30 or 80 ppm) exposure for 8 weeks suppressed the GGT activity in a dose dependent manner in thymus and macrophage. In spleen, significant decrease in the enzyme activity was observed at higher exposure (200 ppm DDT or 80 ppm lindane) levels. Lindane suppressed GGT activity at both 30 or 80 ppm dose levels, while DDT reduced the GGT activity at 200 ppm but not at 100 ppm exposure in lymphocyte. The study indicates the possibility of using GGT as an effective and consistent biochemical marker for immunotoxicity of xenobiotics and other environmental stressors.

Animals↗
PubMed1996

Maternal effects of the short integument mutation on embryo development in Arabidopsis.

Maternal gene products deposited in an animal egg determine the polarity of embryonic axes and regulate embryonic cell-cell communication important for morphogenesis. Here we report the first maternal-effect embryo-defective mutation discovered in a plant. Recessive mutations in the SHORT INTEGUMENT (SIN1) gene in Arabidopsis were previously shown to influence ovule development and flowering time. Here we show that a sin1 mutation has a pronounced maternal effect on zygotic embryo development. A homozygous sin1 mutant embryo is normal when nursed by a sin1/+ heterozygous maternal sporophyte. Strikingly, a sin1 or a sin1/+ embryo that is nursed by a sin1 homozygous maternal sporophyte develops morphogenetic defects in the apical-basal and radial axes. The defects resemble those seen in some zygotic-effect embryonic pattern formation mutants. These results imply that in maternal cells the SIN1 gene either codes for or controls the production of a diffusible morphogen necessary for proper zygotic embryogenesis.

Arabidopsis↗
PubMed1996

Interactions of aristololactam beta-D-glucoside with right-handed and left-handed forms of synthetic deoxyribonucleic acid: spectroscopic and thermodynamic study.

The interaction of aristololactam beta-D-glucoside (ADG) with different polymorphic structures of poly(dG-me5dC).poly(dG-me5dC), poly(dG-dC).poly(dG-dC), and poly(dI-dC). poly(dI-dC) has been studied by spectrophotometric, spectrofluorimetric, circular dichroism, UV melting profiles, and thermodynamic analysis. The binding of ADG to B-form duplexes is characterized by the typical bathochromic and hypochromic effects in the absorption spectrum, quenching of steady-state fluorescence intensity, a decrease in fluorescence quantum yield of ADG, an increase in fluorescence polarization anisotropy, an increase of thermal transition temperature, and perturbation in circular dichroic spectrum. Scatchard analysis indicates that ADG binds to the right-handed form of each polymer in a noncooperative manner. Comparative binding parameters determined from absorbance titration by Scatchard analysis, employing the excluded site model, indicate a stronger binding of ADG to the B-form of poly(dG-me5-dC). poly(dG-me5dC) than to the B-form of poly(dG-dC).poly(dG-dC) or poly(dI-dC). poly(dI-dC). Thermodynamic parameters (delta G degree, delta H degree, and delta S degree) obtained by van't Hoff analysis of the data show that the process of binding to all B-form duplexes is exothermic and enthalpy driven as characterized by a favorable negative enthalpy change (delta H degree). The binding is opposed by a negative entropy change (delta S degree) contribution. Conformational changes indicate that the alkaloid converts the left-handed form of poly(dG-dC). poly(dG-dC), and its methylated analogue and high salt form of poly(dI-dC). poly(dI-dC) to a bound right-handed form, while it inhibits both the rate and extent of the B to Z transition. These studies reveal that ADG binds strongly to B-form polymers while it does not bind to polymers of Z-form.

Alkaloids↗
PubMed1996

Thermodynamics of the interactions of sanguinarine with DNA: influence of ionic strength and base composition.

Using a combination of spectrophotometric and spectrofluorimetric techniques, we report the first thermodynamic characterization of sanguinarine binding to a series of natural and synthetic host DNA duplexes over a wide range of temperature and sodium concentration. The binding isotherms fit reasonably well to the neighbour exclusion model. The salt and temperature dependence of the binding constants is used to estimate the thermodynamic parameters involved in the interaction of the alkaloid with DNA. The resulting binding data are found to be sensitive to the ionic strength of the medium, base composition and sequence of base pairs. When the sodium ion concentration is increased from 0.005 M to 0.5 M, the binding free energy changes vary in a range from -8.47 to -7.1 kcal mol-1, which corresponds to a binding constant range from 1.85 x 10(6) to 1.8 x 10(5) M-1 at 20 degrees C. More distinct is the spread in the binding enthalpy changes which range from -6.35 to -2.62 kcal mol-1 corresponding to binding entropy changes from +7.22 to +15.3 cal K-1 mol-1 at 20 degrees C. On the other hand when the GC content of the host DNA duplexes is increased, the binding free energy varies in a range from -7.28 to -8.58 kcal mol-1 with the binding enthalpy changes ranging from -0.46 to -14.31 kcal mol-1, while corresponding binding entropy changes range from +23.3 to -19.56 cal K-1 mol-1 at 20 degrees C. Sanguinarine binding to natural DNAs and homo- and heteropolymers of AT is characterized by negative enthalpy changes and positive entropy changes, while binding to homo- and heteropolymers of GC is reflected by both negative enthalpy changes and entropy changes. Possible molecular contributions towards sign and magnitude of the thermodynamic parameters and their dependence on ionic strength, base composition and sequences, are discussed.

Alkaloids↗
PubMed1996

Rhinovirus stimulation of interleukin-6 in vivo and in vitro. Evidence for nuclear factor kappa B-dependent transcriptional activation.

To further understand the biology of rhinovirus (RV), we determined whether IL-6 was produced during RV infections and characterized the mechanism by which RV stimulates lung cell IL-6 production. In contrast to normals and minimally symptomatic volunteers, IL-6 was detected in the nasal washings from patients who developed colds after RV challenge. RV14 and RV1A, major and minor receptor group RVs, respectively, were potent stimulators of IL-6 protein production in vitro. These effects were associated with significant increases in IL-6 mRNA accumulation and gene transcription. RV was also a potent stimulator of IL-6 promoter-driven luciferase activity. This stimulation was modestly decreased by mutation of the nuclear factor (NF)-IL-6 site and abrogated by mutation of the NF-kappa B site in this promoter. An NF-kappa B-DNA binding activity, mediated by p65, p50, and p52 NF-kappa B moieties, was rapidly induced in RV-infected cells. Activator protein 1-DNA binding was not similarly altered. These studies demonstrate that IL-6 is produced during symptomatic RV infections, that RVs are potent stimulators of IL-6 elaboration, and that RV stimulation IL-6 production is mediated by an NF-kappa B-dependent transcriptional stimulation pathway. IL-6 may play an important role in the pathogenesis of RV infection, and NF-kappa B activation is likely to be an important event in RV-induced pathologies.

Adult↗
PubMed1996

A mutant of Pseudomonas aeruginosa that lacks c-type cytochromes has a functional cyanide-insensitive oxidase.

Using transposon mutagenesis and screening for the loss of the ability to oxidise the artificial electron donor N,N,N',N'-tetramethyl-p-phenylenediamine, we have isolated a mutant of Pseudomonas aeruginosa that lacks all c-type cytochromes. This mutant is unable to grow anaerobically with nitrate as a terminal electron acceptor. Analysis of its respiratory function indicates that the mutant has lost its cytochrome c oxidase-terminated respiratory pathway but the cyanide-insensitive oxidase-terminated branch remains functional. Complementation of the mutant by in vivo cloning led to recovery of the wild-type characteristics. These data are consistent with the idea that the cyanide-insensitive respiratory pathway does not contain haem c and that the pathway's terminal oxidase is a quinol oxidase.

Electron Transport Complex IV↗