Biomedical subjects

R L Warner

Publications and source records attributed to R L Warner.

At least 55 records · Page 3Linked to original sources

PubMed1995

Lung sources and cytokine requirements for in vivo expression of inducible nitric oxide synthase.

Products of inducible nitric oxide synthase (iNOS) are known to be involved in lung injury following intrapulmonary deposition of immunoglobulin G immune complexes (IgG-ICx). In the current studies rat alveolar macrophages stimulated in vitro with murine interferon gamma (IFN-gamma), tumor necrosis factor alpha, interleukin 1 alpha, (IL-1 alpha), lipopolysaccharide (LPS), or IgG-ICx immunostained for iNOS and produced nitrite/nitrate- (NO2-/NO3-) in a dose- and time-dependent manner requiring availability of L-arginine. Under the same conditions, IL-4 and IL-10 reduced NO2-/NO3- generation. Type II alveolar epithelial cells, which were obtained from normal rat lungs and stimulated in vitro with IgG-ICx, LPS, or IFN-gamma, also immunostained for iNOS and generated NO2-/NO3-. Special techniques of bronchoalveolar lavage (BAL) were used to retrieve alveolar macrophages and type II alveolar epithelial cells. Under these conditions, intrapulmonary deposition of LPS yielded BAL fluids containing increased amounts of NO2-/NO3- and macrophages that spontaneously released NO2-/NO3- and stained for iNOS. After intrapulmonary deposition of IgG both macrophages as well as type II cells (retrieved by BAL) spontaneously produced NO2-/NO3- and both cell types immunostained for iNOS (approximately 20% of all type II cells and 35% of all alveolar macrophages). Using dual fluorescence staining for cell identification, frozen sections of lung tissue after IgG immune complex deposition revealed iNOS in both alveolar macrophages and type II cells. Finally, in the immune complex model of alveolitis, the appearance of iNOS in macrophages as well as macrophage production in vitro of NO2-/NO3- was dependent on the in vivo availability of tumor necrosis factor alpha, IL-1, and IFN-gamma. These studies suggest a dual cell source for nitric oxide in inflamed lungs and the requirements for iNOS of several cytokines.

Amino Acid Oxidoreductases↗

PubMed1994

Transcranial electrostimulation effects on rat opioid and neurotransmitter levels.

A specific form of Transcranial Electrostimulation Treatment (TCET) has been shown to induce analgesia, alleviate symptoms of opiate withdrawal and alter nociceptive responses in neurons in the midbrain and hypothalamus of rats. TCET consists of a 10Hz, charge balanced, 10 mu A current passed for 30 minutes between electrodes placed in the ears. Both serotonin (5HT) and endogenous opioids have been strongly implicated in TCET responses. This study directly measured brain levels of several neurotransmitters and their metabolites in anesthetized rats stimulated with either 10 mu A TCET or 0 mu A (Sham). Neurotransmitters measured in selected homogenized brain areas by high performance liquid chromatography were 5HT and its metabolite, 5-hydroxyindolacetic acid (5HIAA); norepinephrine (NE) and its metabolite, 3-methoxy-4-hydroxyphenethyleneglycol (MHPG); and dopamine (DA). Levels of NE and DA were significantly higher in the hypothalamic region of TCET rats than of control rats. The midbrains of TCET rats contained significantly elevated levels of DA, MHPG, 5HT and 5HIAA. In the hindbrain no significant differences were observed. Thus, TCET appears to cause an increase in the synthesis or release of 5HT, DA and NE in the midbrain and DA and 5HT in the hypothalamus. In a separate experiment, beta-endorphin-like immunoreactivity was measured in blood plasma taken from rats at intervals before, during and after a 30 minute TCET treatment, but no demonstrable TCET effect was observed. The lack of change in serum endorphin levels suggests that TCET-induced opioid activity may be confined to the central nervous system, a reasonable theory because the current passes only through the head.

Animals↗

PubMed1994

In vitro effects of vomitoxin (deoxynivalenol) on T-cell interleukin production and IgA secretion.

The exposure of lymphocyte cultures to vomitoxin was used to determine possible mechanisms by which this naturally occurring toxin induces serum immunoglobulin A (IgA) elevation and IgA nephropathy in the mouse. Vomitoxin exposure within the range of 10 to 1000 ng/ml inhibited DNA synthesis, protein synthesis as well as IgA, IgG and IgM production in lymphocyte cultures prepared from the Peyer's patch (PP) and spleen. When purified B cells were cultured in the presence of vomitoxin, inhibition of IgA, IgG and IgM production was similarly observed. However, on 24-hr pulsed co-exposure to vomitoxin and the mitogen concanavalin A (ConA), CD4+/CD8+ cells were capable of inducing a three- to five-fold increase in production of IgA, but not IgG and IgM by cocultured B cells when compared with B cells cocultured with control T cells exposed to the mitogen only. When pulsed for 48 hr with ConA and toxin, CD4+ cells were similarly capable of causing a significant increase in IgA production by B cells. 48-hr pulsed exposure of CD4+ cells to ConA and vomitoxin resulted in significantly increased production of the T helper cytokines IL-4, IL-5 and IL-6 after 5 additional days of culture, compared with ConA-stimulated CD4+ cells alone. These results suggest that vomitoxin was capable of enhancing CD4(+)-mediated help for IgA production by B cells and that this could possibly be mediated by way of increased cytokine production.

Animals↗

PubMed1992

Root respiration associated with ammonium and nitrate absorption and assimilation by barley.

We examined nitrate assimilation and root gas fluxes in a wild-type barley (Hordeum vulgare L. cv Steptoe), a mutant (nar1a) deficient in NADH nitrate reductase, and a mutant (nar1a;nar7w) deficient in both NADH and NAD(P)H nitrate reductases. Estimates of in vivo nitrate assimilation from excised roots and whole plants indicated that the nar1a mutation influences assimilation only in the shoot and that exposure to NO(3) (-) induced shoot nitrate reduction more slowly than root nitrate reduction in all three genotypes. When plants that had been deprived of nitrogen for several days were exposed to ammonium, root carbon dioxide evolution and oxygen consumption increased markedly, but respiratory quotient-the ratio of carbon dioxide evolved to oxygen consumed-did not change. A shift from ammonium to nitrate nutrition stimulated root carbon dioxide evolution slightly and inhibited oxygen consumption in the wild type and nar1a mutant, but had negligible effects on root gas fluxes in the nar1a;nar7w mutant. These results indicate that, under NH(4) (+) nutrition, 14% of root carbon catabolism is coupled to NH(4) (+) absorption and assimilation and that, under NO(3) (-) nutrition, 5% of root carbon catabolism is coupled to NO(3) (-) absorption, 15% to NO(3) (-) assimilation, and 3% to NH(4) (+) assimilation. The additional energy requirements of NO(3) (-) assimilation appear to diminish root mitochondrial electron transport. Thus, the energy requirements of NH(4) (+) and NO(3) (-) absorption and assimilation constitute a significant portion of root respiration.

Journal Article↗

PubMed1991

Characterization and sequence of a novel nitrate reductase from barley.

Barley (Hordeum vulgare L.) has both NADH-specific and NAD(P)H-bispecific nitrate reductases. Genomic and cDNA clones of the NADH nitrate reductase have been sequenced. In this study, a genomic clone (pMJ4.1) of a second type of nitrate reductase was isolated from barley by homology to a partial-length NADH nitrate reductase cDNA and the sequence determined. The open reading frame encodes a polypeptide of 891 amino acids and its interrupted by two small introns. The deduced amino acid sequence has 70% identity to the barley NADH-specific nitrate reductase. The non-coding regions of the pMJ4.1 gene have low homology (ca. 40%) to the corresponding regions of the NADH nitrate reductase gene. Expression of the pMJ4.1 nitrate reductase gene is induced by nitrate in root tissues which corresponds to the induction of NAD(P)H nitrate reductase activity. The pMJ4.1 nitrate reductase gene is sufficiently different from all previously reported higher plant nitrate reductase genes to suggest that it encodes the barley NAD(P)H-bispecific nitrate reductase.

Amino Acid Sequence↗

PubMed1991

Triple synchronous primary lung carcinomas treated with simultaneous resection.

The fifth case of surgical treatment for three primary bronchogenic lung cancers is presented. This is the second successful surgical case in which this occurred bilaterally and the first case in which simultaneous resection via median sternotomy for three separate primary bronchogenic carcinomas was performed. We strongly advocate this approach when feasible.

Carcinoma, Bronchogenic↗

PubMed1990

Elevated membrane IgA+ and CD4+ (T helper) populations in murine Peyer's patch and splenic lymphocytes during dietary administration of the trichothecene vomitoxin (deoxynivalenol).

Recent investigations indicate that dietary exposure to the trichothecene vomitoxin increases total and antigen-specific serum immunoglobulin A (IgA) and glomerular IgA accumulation in mice. In this study, the effects of 25 ppm dietary vomitoxin on the histological and lymphocytic profile of component immune organs in the mucosal lymphocyte migratory pathway were evaluated in the B6C3F1 mouse. Vomitoxin administration resulted in marked stimulation of the size and frequency of germinal centres in Peyer's patches, mesenteric lymph nodes and the spleen. A slight increase in the percentage of B cells in the Peyer's patch was observed, although vomitoxin treatment had no effect on the percentage of B cells in the spleen. The percentage of IgA+ cells in Peyer's patches and spleen were approximately twice that of controls at 4, 8 and 12 wk of vomitoxin exposure whereas the percentage of IgG+ cells decreased in these two organs. Exposure to vomitoxin increased the percentage of T cells in Peyer's patches and the spleen. The percentage of CD4+ cells (T helper subset) increased slightly in Peyer's patches and more markedly (30-50%) in the spleen following vomitoxin treatment. Contrastingly, there was only a slight increase in the percentage of CD8+ cells (T cytotoxic/suppressor subset) in the spleens of vomitoxin-treated mice in comparison with controls, and no effect in Peyer's patches. The relative effects of vomitoxin on these two T cells populations was also reflected in increased CD4+: CD8+ ratios in Peyer's patches and spleen. These results are consistent with the hypothesis that dietary vomitoxin modulates normal regulation of the IgA response at the Peyer's patch level and that this is manifested in an altered lymphocyte distribution pattern in both the mucosal and systemic compartment. Notably increased levels of IgA+ and CD4+ cells are indicative of IgA-producing progenitors and T helper subsets, respectively, that in tandem could favour IgA hyperproduction and elevated IgA in serum.

Animals↗

PubMed1990

Effect of dietary administration of the trichothecene vomitoxin (deoxynivalenol) on IgA and IgG secretion by Peyer's patch and splenic lymphocytes.

Prolonged dietary exposure of mice to the trichothecene vomitoxin induces abnormally high levels of serum IgA and kidney mesangial IgA accumulation in a manner that is highly analogous to the human glomerulonephritis IgA nephropathy. In this study, the capacity of Peyer's patch and splenic lymphocytes to produce IgA and IgG were compared in B6C3F1 mice that were fed diets with and without 25 ppm vomitoxin for up to 12 wk. Serum IgA increased 2-, 4- and 8-fold after 4, 8 and 12 wk, respectively, of vomitoxin exposure and it became the primary serum isotype, whereas serum IgG was unaffected. On termination of the experiment there were increased numbers of IgA-secreting cells in Peyer's patches after 8 wk of toxin exposure and in the spleen after 4, 8 and 12 wk of toxin exposure. There were also increased numbers of IgG-secreting cells in Peyer's patches on termination of the experiment at 4, 8 and 12 wk but no effects was observed in the spleen. Supernatant IgA and IgA-secreting cell numbers were also markedly elevated in lymphocyte cultures obtained from Peyer's patches and, to a lesser extent, from spleens of treated mice compared with controls. Based on output of treated mice relative to corresponding controls, IgA secretion was greatest in concanavalin-A-stimulated and unstimulated Peyer's patch cultures. Enhanced IgG secretion and IgG-secreting cells were also observed in mitogen-stimulated and unstimulated Peyer's patch lymphocyte cultures of treated relative to control mice, but differences in splenocyte cultures were negligible. Based on total Ig output, IgA production was 8- to 20-fold greater than IgG production in both control and treatment Peyer's patch cultures. In contrast, vomitoxin treatment caused a shift from primarily IgG production in lipopolysaccharide-stimulated spleen cultures to equivalent IgA production. These data provide in vitro evidence that ingestion of vomitoxin promotes terminal differentiation of IgA-secreting progenitors in the Peyer's patch and, to a lesser extent, in the spleen. These functional changes are consistent with the shift from IgG to IgA as the primary serum isotype.

Animals↗

PubMed1990

Altered serum immunoglobulin response to model intestinal antigens during dietary exposure to vomitoxin (deoxynivalenol).

The effect of dietary vomitoxin on the serum IgA and IgG responses to two model intestinal antigens, casein and cholera toxin (CT), were assessed in 4 experimental groups: (1) mice fed casein-based diet, (2) mice fed casein-based diet containing 25 ppm vomitoxin, (3) mice fed casein-based diet and immunized with CT, and (4) mice fed casein-based diet containing 25 ppm vomitoxin and immunized with CT. Unimmunized and CT-immunized mice that were fed vomitoxin exhibited increased levels of total serum IgA relative to matched control animals fed the standard diet. Relative concentrations of casein-specific IgA were greater in both unimmunized mice and CT-immunized mice fed standard diet with vomitoxin than in matched controls fed standard diet only. CT-specific serum IgA in CT-immunized mice was not affected by vomitoxin feeding, but relative levels of CT-specific IgA were higher in unimmunized mice fed vomitoxin than in unimmunized mice fed standard diet. Both casein- and CT-specific serum IgG were depressed in mice fed vomitoxin. Significant differences in total, casein-specific and CT-specific IgA within the intestinal contents were not observed between CT-immunized mice fed vomitoxin and those fed the control diet. The results suggest that vomitoxin altered regulation of the normal immunoglobulin response to intestinal antigens and that this was manifested in the systemic compartment.

Animals↗

PubMed1989

Dysregulation of IgA production and IgA nephropathy induced by the trichothecene vomitoxin.

The effect of dietary exposure to vomitoxin on serum immunoglobulin A (IgA) was evaluated in the B6C3F1 mouse. Levels of serum IgA were elevated maximally in mice fed 25 ppm vomitoxin in comparison with levels in mice fed 2, 10 or 50 ppm vomitoxin. Significant increases were detectable after as few as 4 wk in mice fed 25 ppm vomitoxin, and IgA levels were increased more than 17-fold after 24 wk of toxin exposure. Serum IgA also exhibited a marked shift from primarily monomeric IgA to primarily polymeric IgA during vomitoxin treatment. Serum IgG and IgM decreased in treated mice, suggesting that the effect was isotype-specific. Elevated serum IgA was not observed in mice when control diet was fed at levels equivalent to those consumed by vomitoxin-treated mice, which exhibited feed refusal. IgA production was significantly increased in both spontaneous and mitogen-stimulated splenocyte cultures from mice exposed to vomitoxin in comparison with cultures prepared from ad lib. or feed-restricted controls. Immunofluorescence staining revealed marked accumulation of mesangial IgA and electron microscopy showed electron-dense deposits in the glomeruli of vomitoxin-treated mice but not in those of controls. Dysregulation of IgA production and accumulation of glomerular IgA as observed in this study were highly analogous to the characteristics of human IgA nephropathy, the most common form of glomerulonephritis worldwide.

Animals↗

PubMed1989

Traumatic pancreatitis and pseudocyst in children: current management.

Twenty years ago, we reviewed the pediatric surgical experience with traumatic pancreatitis and pseudocysts at a large children's hospital. That series encompassed 13 years, during which time 23 cases were found. Six of these had pseudocysts, five of which were managed by external catheter drainage and one by excision. The present series spans 10 years and consists of nine cases of post-traumatic pediatric pancreatitis, seven of which progressed to pseudocysts. Four of these were externally drained, one was excised, and two resolved spontaneously. We have reviewed this recent experience in order to re-evaluate the efficacy of external drainage of pseudocysts in pediatric patients. We continue to believe that external drainage is the most effective method of therapy. Internal drainage is usually unnecessary unless the drainage from the ductal disruption does not resolve.

Abdominal Injuries↗

PubMed1989

Reduced Accumulation of ABA during Water Stress in a Molybdenum Cofactor Mutant of Barley.

A barley (Hordeum vulgare L.) mutant (Az34) has been identified with low basal levels of abscisic acid (ABA) and with reduced capacity for producing ABA in response to water stress. The mutation is in a gene controlling the molybdenum cofactor resulting in a pleiotropic deficiency in at least three molybdoenzymes, nitrate reductase, xanthine dehydrogenase, and aldehyde oxidase. The mutant was found to lack aldehyde oxidase activity with several substrates including: (a) ABA aldehyde, a putative precursor of ABA; (b) an acetylenic analog of ABA aldehyde; and (c) heptaldehyde. Elevating the growth temperature from 18 to 26 degrees C caused mutant leaves to wilt and brown. Desiccation of mutant leaves was prevented by applying ABA. These results indicate that ABA biosynthesis at some developmental stages is dependent upon a molybdoenzyme which may be an aldehyde oxidase.

Journal Article↗

PubMed1989

Oxygen and carbon dioxide fluxes from barley shoots depend on nitrate assimilation.

A custom oxygen analyzer in conjunction with an infrared carbon dioxide analyzer and humidity sensors permitted simultaneous measurements of oxygen, carbon dioxide, and water vapor fluxes from the shoots of intact barley plants (Hordeum vulgare L. cv Steptoe). The oxygen analyzer is based on a calciazirconium sensor and can resolve concentration differences to within 2 microliters per liter against the normal background of 210,000 microliters per liter. In wild-type plants receiving ammonium as their sole nitrogen source or in nitrate reductase-deficient mutants, photosynthetic and respiratory fluxes of oxygen equaled those of carbon dioxide. By contrast, wild-type plants exposed to nitrate had unequal oxygen and carbon dioxide fluxes: oxygen evolution at high light exceeded carbon dioxide consumption by 26% and carbon dioxide evolution in the dark exceeded oxygen consumption by 25%. These results indicate that a substantial portion of photosynthetic electron transport or respiration generates reductant for nitrate assimilation rather than for carbon fixation or mitochondrial electron transport.

Journal Article↗

PubMed1989

Nitrate transport is independent of NADH and NAD(P)H nitrate reductases in barley seedlings.

Barley (Hordeum vulgare L.) has NADH-specific and NAD(P)H-bispecific nitrate reductase isozymes. Four isogenic lines with different nitrate reductase isozyme combinations were used to determine the role of NADH and NAD(P)H nitrate reductases on nitrate transport and assimilation in barley seedlings. Both nitrate reductase isozymes were induced by nitrate and were required for maximum nitrate assimilation in barley seedlings. Genotypes lacking the NADH isozyme (Az12) or the NAD(P)H isozyme (Az70) assimilated 65 or 85%, respectively, as much nitrate as the wild type. Nitrate assimilation by genotype (Az12;Az70) which is deficient in both nitrate reductases, was only 13% of the wild type indicating that the NADH and NAD(P)H nitrate reductase isozymes are responsible for most of the nitrate reduction in barley seedlings. For all genotypes, nitrate assimilation rates in the dark were about 55% of the rates in light. Hypotheses that nitrate reductase has direct or indirect roles in nitrate uptake were not supported by this study. Induction of nitrate transporters and the kinetics of net nitrate uptake were the same for all four genotypes indicating that neither nitrate reductase isozyme has a direct role in nitrate uptake in barley seedlings.

Biological Transport↗

PubMed1989

The influence of electrical variables on analgesia produced by low current transcranial electrostimulation of rats.

Pulsed low current transcranial electrostimulation (TE) has been shown to induce analgesia in rats as measured by the wet tail flick test. This study investigates the effect of varying stimulus frequency, pulse width, charge balance and polarity, as well as the influence of electrode placement and time of day at which stimulus occurred. A biphasic, charge balanced waveform with a first phase duration of 2 msec, current 10 microA and repetition rate 10 Hz was found to induce maximum tail flick latency changes. The effects of morning or nighttime stimulation were statistically indistinguishable, as were the differences between monophasic and biphasic stimulation. Analgesia was maximized when a positive first phase was delivered into the right ears of the rats, but monolateral stimulation with both electrodes on either the left or the right ear produced no measurable effect. Examination of TE responses in sham and stimulated populations reveals normal response distributions with the stimulated group skewed toward a positive effect.

Analgesia↗

PubMed1987

Biochemical and genetic comparison of two nitrate reductase-deficient pea mutants disturbed in the cofactor.

Two nitrate reductase (NaR)-deficient mutants of pea (Pisum sativum L.), E1 and A300, both disturbed in the molybdenum cofactor function and isolated, respectively, from cv Rondo and cv Juneau, were tested for allelism and were compared in biochemical and growth characteristics. The F1 plants of the cross E1 X A300 possessed NaR and xanthine dehydrogenase (XDH) activities comparable to those of the wild types, indicating that these mutants belong to different complementation groups, representing two different loci. Therefore, mutant E1 represents, besides mutant A300 and the allelic mutants A317 and A334, a third locus governing NaR and is assigned the gene destignation nar 3. In comparison with the wild types, cytochrome c reductase activity was increased in both mutants. The mutants had different cytochrome c reductase distribution patterns, indicating that mutant A300 could be disturbed in the ability to dimerize NaR apoprotein monomers, and mutant E1 in the catalytic function of the molybdenum cofactor. In growth characteristics studied, A300 did not differ from the wild types, whereas fully grown leaves of mutant E1 became necrotic in soil and in liquid media containing nitrate.

Coenzymes↗

PubMed1983

Nitrate reductase-deficient mutants in barley : immunoelectrophoretic characterization.

Nitrate reductase-deficient barley (Hordeum vulgare L.) mutants were assayed for the presence of a functional molybdenum cofactor determined from the activity of the molybdoenzyme, xanthine dehydrogenase, and for nitrate reductase-associated activities. Rocket immunoelectrophoresis was used to detect nitrate reductase cross-reacting material in the mutants. The cross-reacting material levels of the mutants ranged from 8 to 136% of the wild type and were correlated with their nitrate reductase-associated activities, except for nar 1c, which lacked all associated nitrate reductase activities but had 38% of the wild-type cross-reacting material. The cross-reacting material of two nar 1 mutants, as well as nar 2a, Xno 18, Xno 19, and Xno 29, exhibited rocket immunoprecipitates that were similar to the wild-type enzyme indicating structural homology between the mutant and wild-type nitrate reductase proteins. The cross-reacting materials of the seven remaining nar 1 alleles formed rockets only in the presence of purified wild-type nitrate reductase, suggesting structural modifications of the mutant cross-reacting materials. All nar 1 alleles and Xno 29 had xanthine dehydrogenase activity indicating the presence of functional molybdenum cofactors. These results suggest that nar 1 is the structural gene for nitrate reductase. Mutants nar 2a, Xno 18, and Xno 19 lacked xanthine dehydrogenase activity and are considered to be molybdenum cofactor deficient mutants. Cross-reacting material was not detected in uninduced wild-type or mutant extracts, suggesting that nitrate reductase is synthesized de novo in response to nitrate.

Journal Article↗

PubMed1983

Synthesis and degradation of barley nitrate reductase.

Nitrate and light are known to modulate barley (Hordeum vulgare L.) nitrate reductase activity. The objective of this investigation was to determine whether barley nitrate reductase is regulated by enzyme synthesis and degradation or by an activation-inactivation mechanism. Barley seedling nitrate reductase protein (cross-reacting material) was determined by rocket immunoelectrophoresis and a qualitative immunochemical technique (western blot) during the induction and decay of nitrate reductase activity. Nitrate reductase cross-reacting material was not detected in root or shoot extracts from seedlings grown without nitrate. Low levels of nitrate reductase activity and cross-reacting material were observed in leaf extracts from plants grown on nitrate in the dark. Upon nitrate induction or transfer of nitrate-grown etiolated plants to the light, increases in nitrate reductase activity were positively correlated with increases in immunological cross-reactivity. Root and shoot nitrate reductase activity and cross-reacting material decreased when nitrate-induced seedlings were transferred to a nitrate-free nutrient solution or from light to darkness. These results indicate that barley nitrate reductase levels are regulated by de novo synthesis and protein degradation.

Journal Article↗