Low-level hyperbaric heliox does not affect drug-induced seizure latency in mice.
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Biomedical subjects
Publications and source records attributed to B L Jones.
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The behavioral manifestations of acute ethanol intoxication resemble those of benzodiazepines, barbiturates and general anesthetics. This has led to speculation that these drugs share common mechanisms or sites of actions within the brain. The discovery of a specific benzodiazepine receptor site, and the subsequent development of selective receptor antagonist and inverse agonist drugs, provides a framework to test the involvement of the benzodiazepine receptor complex in mediating ethanol's behavioral effects. The partial inverse agonist Ro15-4513, an analog of the benzodiazepine receptor antagonist Ro15-1788 (flumazenil), has been reported to block or reduce some of ethanol's acute effects in rodents by a benzodiazepine receptor-mediated action. There has been some controversy over whether the "antialcohol" effect of Ro15-4513 is a unique property of this compound or is shared by other benzodiazepine antagonists with inverse agonist activity. We have studied the effects of Ro15-4513 and other benzodiazepine receptor antagonists on acute ethanol intoxication in mice and have obtained evidence that 1) Ro15-4513 differentially affects acute effects of ethanol, 2) an "antialcohol" property is not a general feature of all benzodiazepine antagonists and inverse agonists, and 3) "antialcohol" activity may not be unique to Ro15-4513.
The hydrolytic specificity of a 30 kilodalton cysteine proteinase purified from germinated barley (Hordeum vulgare L. cv Morex) was investigated using high performance liquid chromatography to characterize its hydrolysis of two small barley seed proteins, the alpha- and beta-hordothionins. The reduced and pyridylethylated thionins were rapidly cleaved, resulting in the production of a limited number of peptides. Peptide bonds Gly9-Arg10, Cys 16-Arg17, Cys25-Ala26, and Thr34-Ser35 were most susceptible to hydrolysis, the peptide bonds Arg5-Ser6, Arg19-Gly20 in both thionins and Lys38-Cys39 in beta-hordothionin and Cys29-Arg30 of alpha-hordothionin being broken at much slower rates. The hydrolysis patterns were highly reproducible from assay to assay and with various enzyme preparations. The specificity was apparently defined by the amino acids in the P(2) position, not those immediately adjacent to the susceptible bonds. The P(2) amino acid residues of the released peptides were always either leucine, valine, tyrosine, or pyridylethylcysteine. From these observations and from the rates of release of the various peptides, it appears that the barley 30 kilodalton endoproteinase has an S2 subsite that preferentially binds the leucine side chain: i.e. for hydrolyzing the peptide bond P(1)-P(1)' in the general sequence NH(2)-P(2)-P(1)-P(1)'-COOH, the enzyme is selective for leucine and, to a lesser extent, valine and tyrosine at position P(2). The barley proteinase thus resembles two other cysteine proteinases, papain and Streptococcal proteinase, in its specificity.
Offsetting ethanol-induced hypothermia in five inbred strains of mice changed ethanol sensitivity within strains and markedly reduced differences between strains in brain sensitivity to hypnotic ethanol doses. The present study extended this work to mice selectively bred for sensitivity and resistance to ethanol-induced loss of righting reflex (LORR) and hypothermia. In all experiments LORR duration and ethanol concentrations at return of righting reflex were measured after i.p. hypnotic ethanol doses and exposure to 22 or 34 degrees C. In experiment 1, C57BL/6J, A/HeJ, 129/J, LS/lbg and SS/lbg mice were given 4.2 g/kg ethanol. In experiment 2, the same mouse genotypes were tested with different ethanol doses (2.5-4.9 g/kg) selected to produce an equivalent degree of impairment (60 min LORR duration). In experiment 3, HOT and COLD lines of mice were given 4.0 g/kg ethanol. In agreement with previous work, offsetting hypothermia reduced differences between genotypes in ethanol sensitivity. Comparisons within genotypes indicated that ethanol sensitivity in C57, A/He, SS, HOT and COLD mice increased as body temperature increased. In contrast, ethanol sensitivity in 129 and LS mice decreased as body temperature increased. These results extend previous findings indicating that body temperature during intoxication contributes to differences between genotypes in ethanol sensitivity. The present findings also suggest that there are qualitative differences in the effects of temperature on ethanol sensitivity within genotypes.
The effect of ambient and body temperature on ethanol lethality in inbred strains and selected lines of mice was investigated. C57BL/6J, 129/J, LS/Ibg and SS/Ibg mice were exposed to 23 or 34 degrees C following IP injection of lethal ethanol doses (8.2, 6.0, 6.5 or 7.0 g/kg ethanol, respectively). All mice exposed to 23 degrees C during intoxication became markedly hypothermic, with mean body temperatures dropping to lows of 27.9, 30.3, 33.0 and 33.3 degrees C in C57, LS, SS and 129 animals, respectively. Compared to the 23 degrees C groups, exposure to 34 degrees C offset the ethanol-induced hypothermia and significantly increased percent mortality in all four mouse genotypes. Exposure to 34 degrees C increased mortality at 24 hours postinjection from 15% to 95% in SS mice, from 37.5% to 100% in 129 mice and from 50% to 100% in LS and C57 mice. Blood ethanol data suggest that the present results cannot be explained by temperature-related changes in ethanol elimination. These results provide further evidence that body temperature during intoxication can have major effects on mortality rates in mice.
In order to identify and quantify tasks performed by pharmacists and technicians in the Newborn Center Satellite Pharmacy, a workload assessment using a time ladder log was performed. Two pharmacists and one technician documented their daily activities in 2-minute increments for 1 month. Tasks were divided into three categories: patient days, calendar days, and variable activities. The average time to perform each task was determined. Standard times for each category were recorded onto a monthly workload report, which was used to calculate full-time equivalents (FTEs) required to perform all tasks. The number of FTEs increased seasonally, as did the number of patient days. Therefore, this workload reporting system can be used to identify peak workload time periods and to justify staffing requirements.
Dynamic configurational changes in the rectus femoris muscle were examined using a real time ultrasound sector scanner in ten normal subjects. The angle of passive knee flexion was varied as transverse ultrasound scanning was performed at the mid-transverse thigh. Configurational changes in the rectus femoris were computed from a traced outline of the muscle and the geometric center of the mass was calculated at all degrees of knee flexion. The geometric center of the mass varied with knee position. The anterioposterior dimensions and cross sectional areas of the rectus femoris and vastus intermedius remained constant, however, despite changes in knee position. The pattern of change observed was reproducible and reflects consistent changes in muscle configuration. The technique and instrumentation should have value for non-invasive dynamic and static observation of individual muscles.
We assessed postnatal growth, neurodevelopmental outcome, and occurrence of respiratory illnesses in 46 infants of very low birth weight who were enrolled in a randomized, controlled, bicenter clinical trial of human surfactant treatment for respiratory distress syndrome. No long-term adverse effects of human surfactant treatment were detected between control and human surfactant-treated infants with respect to growth, neurologic, or developmental outcome. Infants with chronic lung disease, regardless of treatment group, had poorer growth and were more likely to have neurodevelopmental abnormalities at 12 to 24 months of age.
A proteinase was purified from germinated barley (green malt from Hordeum vulgare L. cv Morex) by acidic extraction, ammonium sulfate fractionation and successive chromatographies on CM-cellulose, hemoglobin sepharose, Sephadex G-75 and organomercurial agarose columns. The overall purification and final recovery were 290-fold and 7.5%, respectively. The purified enzyme was homogeneous on analytical gel electrophoresis, yielding a single protein associated with protease activity. An apparent molecular weight of about 20 kilodaltons was estimated for the native enzyme from gel filtration. SDS-gel electrophoresis revealed a single polypeptide of about 30 kilodaltons. The optimum pH for the hydrolysis of hemoglobin was around 3.8. The enzyme was strongly inhibited by leupeptin but was insensitive to phenylmethylsulfonyl fluoride, indicating that it was a cysteine proteinase. It hydrolyzed several large proteins from various origins. The ability of the enzyme to digest barley storage proteins in vitro was examined using SDS-gel electrophoresis. The hydrolysis patterns obtained showed that the enzyme rapidly hydrolyzed the large hordein polypeptides into relatively small fragments. The results of this study suggest that this 30 kilodalton enzyme is one of the predominant cysteine proteinases secreted into the starchy endosperm during barley germination and that it plays a major role in the mobilization of storage proteins.
To determine whether the Triticum urartu genome is more closely related to the A or B genome of the polyploid wheats, the amino acid sequence of its purothionin was compared to the amino acid sequences of the purothionins in Triticum monococcum, Triticum turgidum, and Triticum aestivum. The residue sequence of the purothionin from T. urartu differs by five and six amino acid substitutions respectively from the alpha 1 and alpha 2 forms coded for by genes in the B and D genomes, and is identical to the beta form specified by a gene in the A genome. Therefore, the T. urartu purothionin is either coded by a gene in the A genome or a chromosome set highly homologous to it. The results demonstrate that at least a portion of the T. urartu and T. monococcum genomes is homologous and probably identical. A variety of other studies have also shown that T. urartu is very closely related to T. monococcum and, in all likelihood, also possesses the A genome. Therefore, it could be argued that either T. urartu and T. monococcum are the same species or that T. urartu rather than T. monococcum is the source of the A genome in T. turgidum and T. aestivum. Except for Johnson's results, our data and that of others suggest a revised origin of polyploid wheats. Specifically, the list of six putative B genome donor species is reduced to five, all members of the Sitopsis section of the genus Aegilops.
The solution structure of the 45-residue plant protein, alpha 1-purothionin, is investigated by nuclear magnetic resonance (n.m.r.) spectroscopy. Using a combination of two-dimensional n.m.r. techniques to demonstrate through-bond and through-space (less than 5 A) connectivities, the 1H n.m.r. spectrum of alpha 1-purothionin is assigned in a sequential manner. The secondary structure elements are then delineated on the basis of a qualitative interpretation of short-range nuclear Overhauser effects (NOE) involving the NH, C alpha H and C beta H protons. There are two helices extending from residues 10 to 19 and 23 to 28, two short beta-strands from residues 3 to 5 and 31 to 34 which form a mini anti-parallel beta-sheet, and five turns. In addition, a number of long-range NOE connectivities are assigned and a low resolution tertiary structure is proposed.
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Exogenous cyclic guanosine-3',5'-monophosphate (cGMP) inhibited the growth of Rhizobium japonicum at less than 100 microM. Other nucleotides, including cyclic AMP, cyclic IMP, and cyclic CMP, had no inhibitory effect even at higher concentrations nor was the inhibition by cGMP reversed by cyclic AMP. The inhibitory effect was independent of the carbon and nitrogen source(s) used. cGMP did not inhibit the growth of any other species of bacterium tested, including several fast-growing Rhizobium species. The kinetics of growth inhibition are multiphasic, with no apparent effect for several hours after addition, followed by a period of total inhibition. Subsequently, growth resumed at a slower rate. Resumption of growth was not due to destruction of the nucleotide. Studies of the intracellular cGMP concentration did not reveal significant changes in cells grown under aerobic or microaerobic conditions. No effect of cGMP on the derepression of respiratory nitrate reductase was observed.
We describe a noninvasive quantitative way of measuring muscle with a specially-designed digital ultrasound scanner. Reliability and reproducibility of echo amplitude were determined in 16 normal male volunteers--10 runners and 6 non-runners. Echo amplitudes were recorded from the quadriceps femoris muscle. The muscle was scanned at mid-thigh in a relaxed state with 18 degrees flexion and in an isometrically contracted state with the leg in full extension. Echo amplitudes obtained in the non-runner group were: (1) mean value of the muscle in the relaxed state (174.9 +/- 30.8) and (2) mean value of the muscle in the contracted state (121.8 +/- 31.4). The difference in these values (53.1) was significant at p less than 0.05. Echo amplitudes for runners were: (1) mean value of the muscle in the relaxed state (146.6 +/- 39.2) and (2) mean value of the muscle in the contracted state (107.1 +/- 30.1). The difference in these values (39.5) was significant at p less than -0.01. The data indicate that: values of echo amplitudes are reproducible; there is a statistically significant difference in the mean amplitudes of muscle in a contracted and relaxed state; and while the difference between runner and non-runner muscle is not statistically significant (p greater than 0.1), there is a trend toward significance. Because ultrasound amplitude is a function of tissue structure, this technique may provide a noninvasive method for quantifying muscle collagen and fat.
Purothionins were extracted and purified from the diploid wheat Triticum monococcum. Two proteins were obtained, one of which was present in only very small amounts. The major purothionin of T. monococcum was sequenced and it had an amino acid sequence identical with that of the beta-purothionin of Triticum aestivum (hexaploid bread wheat). It is known that T. monococcum contains the wheat A genome, so the structural gene coding for the beta-purothionin must comprise a part of the A genome. There have been no observable (as amino acid replacements) changes in the DNA comprising either the beta-purothionin gene of T. aestivum or the purothionin gene of T. monococcum, since T. monococcum (or its wild equivalent, Triticum boeoticum) hybridized with the diploid wheat B genome progenitor and started the evolution from diploid to allohexaploid wheat. All of the investigated characteristics of the purothionin-like protein isolated in small amounts suggested that it was essentially identical in amino acid sequence with the T. monococcum purothionin. It may be a dimerized form of beta-purothionin.
Ambient outdoor concentrations and size distributions of airborne microbial particles were measured approximately weekly for 2 years in a Washington, D.C., suburban area. The study objective was to characterize microbial air quality in the vicinity of a proposed sewage sludge composting facility. During the study, 379 samples were taken at 17 stations, using Andersen microbial samplers. Concentration ranges (in viable particles per cubic meter) were as follows: airborne mesophilic fungi, 0 to 7,220 with a geometric mean of 273; thermophilic fungi, 0 to 193 with a median of 2.1; Aspergillus fumigatus, 0 to 71 with a median of 1.0; aerobic bacteria, 4.2 to 1,640 with a geometric mean of 79; and fecal streptococci, 0 to 5.7 with a median of 0. No fecal coliforms were recovered. The potentially respirable fraction (less than 8 microns) averaged 34% for total bacteria, 56% for mesophilic fungi, 91% for thermophilic fungi, and 95% for A. fumigatus. The specific sampling location was not a major factor affecting microbial particle concentrations or size distributions. Conversely, the time of year was an important determinant of viable particle concentrations for all groups of microorganisms studied. The highest concentrations were observed in summer and fall, with significantly lower levels detected in winter. In general, the microbial data did not correlate with other variables, including weather conditions, measured in this study.
The thionins comprise a group of very basic proteins of Mr approximately 5000 found in the seeds of Gramineae. They each contain 45 amino acid residues arranged along a single polypeptide chain that is constrained by four disulfide bridges. Five thionins of known sequence, from barley and wheat, have been investigated and compared by 1H NMR spectroscopy at 600 MHz. From their spectral characteristics it is concluded that the five proteins have very similar, nonrandom conformations in 2H2O solution. Moreover, on the basis of selective nuclear Overhauser experiments at 300 MHz, features of their secondary and tertiary structures are shown to be similar to those of crambin, a related, hydrophobic protein extracted from seeds of the crucifer Crambe abyssinica. The strong compositional homology of the thionins facilitates the assignment of methyl and aromatic resonances, as only a few residues are replaced and these are at known sites. The substitution of leucine for an isoleucine does not affect significantly the local magnetic environment, suggesting that those isomeric side chains easily accommodate the same spatial constraints. A fast hydrogen-deuterium exchange is observed at pH 6.25, 25 degrees C. This indicates that, although of folded conformation, the thionins are structurally flexible polypeptides that efficiently expose all amides to the solvent.