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Biomedical subjects

D R Nelson

Publications and source records attributed to D R Nelson.

At least 217 records · Page 12Linked to original sources

Abomasal impaction in sheep.

Abomasal engorgement was diagnosed in five Suffolk rams and one Suffolk ewe. All sheep had poor or no appetite for between seven days and five months duration. They were depressed, thin and weak. Rectal temperature and respiration rates were within the normal range. Pulse rate was increased (97.6 +/- 11.55/minute, mean +/- se). A distended abdomen was present and, or, a firm mass was ballotted or palpated in the lower right abdomen. Rumen motility was increased and contractions were strong. Laparotomy revealed enlarged abomasums in four sheep. Enlarged abomasums were found at necropsy in two sheep. Abomasal contents resembled rumen ingesta in character and odour. Removal of ingesta by abomasotomy produced temporary improvement in three rams for six to 12 weeks. All six sheep died or were euthanased.

Abomasum↗

Evidence for long-lived mRNA during fruiting body formation in myxococcus xanthus.

Half-lives of Myxococcus xanthus mRNA were determined by inhibiting RNA polymerase with rifampin and then measuring the rate of [35S]methionine incorporation into protein. Vegetative cells resuspended in clone fruiting liquid culture showed an average mRNA half-life of approximately 3.5 min. Developmental cells (24 or 48 hr) exhibited biphasic decay curves with apparent mRNA half-lives of approximately 3.5 min and 20-30 min. The more-stable mRNA species accounted for about 30% of all mRNA present in 24-hr cells and as much as 40% of all mRNA of 48-hr cells. Analysis of the 35S-labeled proteins by NaDodSO4/polyacrylamide gel electrophoresis showed that only 5-10 major polypeptides are synthesized after 30 min of rifampin treatment. One of these is protein S, the spore surface coat protein, because immunoprecipitation of the 35S-labeled proteins with antisera specific for protein S showed continued synthesis of protein S in the presence of rifampin. The half-life for its mRNA was calculated to be 15-30 min. RNA from vegetative and developing cells was pulse labeled with 32PO4 followed by rifampin treatment. Analysis of the labeled RNA on 3% NaDodSO4/polyacrylamide gels showed 5-10 long-lived mRNA bands in developing cells. These results show that there are several abundant mRNA species synthesized developmentally that are exceptionally long lived. The fact that the majority of the mRNA species show the shorter half-life suggests that developing cells retain the normal mechanism for mRNA degradation.

Bacterial Proteins↗

Pharmacokinetics of chloramphenicol in non-lactating cattle.

The pharmacokinetics and bioavailability of a formulation of chloramphenicol base in propylene glycol were determined following administration of single intravenous (i.v.) and subcutaneous (s.c.) 50 mg/kg doses of chloramphenicol to six non-lactating Holstein cows. Mean serum concentrations of chloramphenicol following i.v. administration of 50 mg/kg declined rapidly from a peak of greater than 100 micrograms/ml to 6.9 micrograms/ml at 12 h after administration. Serum concentrations were not detectable at 24 h after administration. The curve of serum concentrations vs time was characteristic of a two-compartment open model. Mean i.v. data gave a biological half-life of 4.3 h and a volume of distribution of the central compartment of 0.44 l/kg. Serum concentrations of chloramphenicol following s.c. administration of 50 mg/kg rose slowly to a broad peak near 20 micrograms/ml from 3 to 8 h after administration and then declined. These data were also analysed according to a two-compartment open model. The biological half-life was 4.2 h and the volume of distribution of the central compartment was 0.50 l/kg. Significant adverse reactions, including acute collapse, intravascular haemolysis and haemoglobinuria, were observed in cows when dosed i.v. Cows dosed s.c. exhibited local reactions at injection sites. The disadvantages of administration of 50 mg/kg doses of chloramphenicol base in propylene glycol appear to be significant and may outweight the potential advantages of parenteral use of the drug as presently formulated.

Animals↗

Transport and localization of protein S, a spore coat protein, during fruiting body formation by Myxococcus xanthus.

Protein S, the most abundant soluble protein synthesized by Myxococcus xanthus FB during early fruiting body formation, accumulates in the soluble fraction of developing cells, reaching a peak at about 24 h; at late stages of fruiting body formation, protein S is found on the surface of spores (M. Inouye et al. Proc. Natl. Acad. Sci. U.S.A. 76:209-213, 1979). In this study, the transport and localization of protein S were investigated. Cells were fractionated to give osmotic shock, membrane, cytoplasmic, and spore fractions. The various fractions were then analyzed for protein S. Protein S was first detected in the cytoplasmic fraction at about 3 to 6 h of development. However, transport of protein S through the cytoplasmic membrane was not observed until 15 to 18 h of development. Thus, protein S is unusual among translocated proteins in that it accumulates as a soluble cytoplasmic protein before translocation. Biosynthesis of protein S ceased after 48 h; by 72 h, protein S was only found on the surface of spores. Pulse-chase experiments were performed to determine the transport kinetics of protein S. The results showed that in 24-h developing cells, the transport of protein S across the cytoplasmic membrane was rapid, occurring in less than 2 min. However, transport across the outer membrane was slow, requiring 10 to 15 min. Pulses of 15 s with [35S]methionine failed to reveal any short-lived precursor form in immunoprecipitated material separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Isoelectric focusing also failed to detect any precursor form of protein S. Thus, protein S appears to be translocated in the absence of a cleaved signal peptide.

Bacterial Proteins↗

A 13C NMR study of methyl-branched hydrocarbon biosynthesis in the housefly.

Carbon 13 NMR, radiotracer and mass spectrometry studies were performed to confirm that propionate and methylmalonate are incorporated into long chain branched hydrocarbons as the methyl branch unit, to determine whether the branching methyl group was added initially or toward the end of the elongation process, to determine the precursor of methylmalonate, and to examine the metabolism of propionate in the housefly, Musca domestica. The labeled carbons from [3-13C]propionate and [methyl-13C]methylmalonate were incorporated into the branching methyl carbon of the methylalkanes, and into the even numbered carbons in alkanes and alkenes. The labeled carbons from [2-13C]propionate and [1-13C]propionate labeled the tertiary carbon and the carbon adjacent to the teritary carbon, respectively, in the methylalkanes. Mass spectral analysis of the methylalkanes after enrichment from [1-13C]propionate showed that propionate was incorporated during the initial stages of chain synthesis of the terminally branched alkanes. Sodium [2-13C]propionate labeled the odd numbered carbons of both the alkanes and alkenes. These data suggest that propionate is converted to an acetyl derivative, with carbon 3 of propionate converted to the carboxyl carbon of acetate, and carbon 2 of propionate converted to the methyl carbon of acetate. This pathway of propionate metabolism has hitherto only been reported in plants. The labeled carbons from DL-[3,4,5-13C3]valine were incorporated intact (as determined by 13C--13C coupling) into the branching methyl carbon, tertiary carbon, and carbon adjacent to the tertiary carbon, respectively, demonstrating that valine serves as a precursor to the methylmalonate used in branched alkane biosynthesis.

Alkanes↗

Surgical stabilisation of the stifle in cranial cruciate ligament injury in cattle.

Surgical stabilisation of cranial cruciate ligament injury was performed on 23 stifles in 22 dairy cattle ranging in age from 11 months to 14 years. Duration of time from injury until surgery varied from two days to three years. A row of Lembert sutures was placed in both the lateral and medial retinacular tissues. In addition, an imbrication suture was placed from the lateral femorotibial ligament to the lateral or middle patellar ligament in five cattle. External support was used in 15 cattle in the form of a modified Robert Jones/Thomas-Schroeder splint. Thirteen cattle improved markedly, four recovered completely and three improved moderately. Surgery was unsuccessful on seven cases. When surgery was unsuccessful, moderate to severe lesions of degenerative joint disease were usually present.

Animals↗

Immunohistochemical localization of IgG1 and IgG2 in prepartum and lactating bovine mammary tissue.

The differential distributions of IgG1 and IgG2 were determined in prepartum and lactating bovine mammary tissue by indirect immunofluorescence. IgG1 was found predominately within the alveolar epithelial cells and lumens of prepartum tissue whereas IgG2 was largely confined to the stromal area surrounding the alveoli. Both IgG subclasses were confined predominately to the stroma in lactating tissue. Few IgG containing stromal cells were readily distinguished in any of the mammary tissue used in this study.

Animals↗

Estimation of the proportion of non-ammonia-nitrogen reaching the lower gut of the ruminant derived from bacterial and protozoal nitrogen.

1. A method for estimating the proportions of bacterial- and protozoal-N in the total non-ammonia-N reaching the lower gut of the ruminant under steady-state conditions was evaluated. Three trials using two different diets were conducted with a Holstein steer equipped with a rumen cannula and duodenal re-entrant cannulas. 2. An intraruminal primed infusion of (15NH4)2SO4 was administered for 68 h during each trial. Bacteria and protozoa samples were isolated from rumen fluid at approximately 6 h intervals during each infusion period. Total non-ammonia-N was isolated from duodenal digesta samples taken at approximately the same times. All of these samples were analysed for 15N enrichment. A computer program was used to fit equations to the 15N-enrichment curves of bacterial- and protozoal-N. Models of both bacterial- and protozoal-N kinetics consisted of a small pool which equilibrated rapidly with rumen NH3 and a large pool with a fractional turnover rate of 0.045-0.070/h for bacterial-N and 0.056-0.069/h for protozoal-N. 3. Abomasal fluid turnover was estimated by a single injection of polyethylene glycol (molecular weight 4000) into the rumen followed by sampling of rumen fluid and duodenal digesta. 4. Estimates of abomasal fluid turnover, bacterial-N turnover, and protozoal-N turnover were entered into an equation which was adjusted by computer iteration to fit the 15N-enrichment curve of duodenal digesta non-NH3-N generated from each (15NH4)2SO4 infusion period. The computer fit of this equation to the observed results gave estimates of 0:39-0.45 and 0.22-0.41 for the proportion of duodenal non-NH3-N derived from bacterial-N and protozoal-N respectively. 5. This method is potentially useful in estimating microbial protein passage to the lower gut in ruminants. Sampling digesta from the omasum rather than the duodenum would simplify the method and possibly increase the reliability of the estimates.

Abomasum↗

Localization of myxobacterial hemagglutinin in the periplasmic space and on the cell surface of Myxococcus xanthus during developmental aggregation.

During the period of developmental aggregation which precedes fruiting body formation, the bacterium Myxococcus xanthus produces a large amount of a lectin called myxobacterial hemagglutinin (MBHA). Sequential cell washing, osmotic shock, and disruption of developmental cells showed that as much as 90% of the total hemagglutinating activity can be recovered in the wash and shock fractions. Analysis of the wash and shock fluids by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that these fractions are enriched in MBHA. MBHA was detected on the surface of developmental cells but not vegetative cells by immunofluorescent staining procedures. The fluorescence was localized in distinct patches which were usually located at one or both of the cell poles, although patches of fluorescence could also be seen at additional sites as well. The presence of MBHA on the cell surface was also detected by electron microscopy of developmental cells stained with ferritin-conjugated antibody. Most of the cells showed distinct patches of ferritin staining at one or both of the cell poles; nonpolar staining, which was also observed, was always accompanied by membrane protuberances. The amino acid sequence of the NH2 terminus of MBHA was determined and found to be extremely hydrophobic, suggesting that it may function as a nonprocessed signal for transmembrane transport. The site-specific localization of MBHA at the cell poles suggests that it may function in end-to-end cellular interactions during aggregation.

Amino Acid Sequence↗

Adult palatal expansion prostheses.

The technical steps in the construction of two types of adult palatal expanders have been described. These prostheses are complex and require a special level of coordination between the professional team and the dental laboratory. The teamwork involved in the restoration of major maxillary defects contributes to the success of the prosthesis and makes the efforts involved worthwhile.

Adult↗

Incorporation of substrate cell lipid A components into the lipopolysaccharide of intraperiplasmically grown Bdellovibrio bacteriovorus.

The composition of Bdellovibrio bacteriovorus lipopolysaccharide (LPS) was determined for cells grown axenically and intraperiplasmically on Escherichia coli or Pseudomonas putida. The LPS of axenically grown bdellovibrios contained glucose and fucosamine as the only detectable neutral sugar and amino sugar, and nonadecenoic acid (19:1) as the predominant fatty acid. Additional fatty acids, heptose, ketodeoxyoctoic acid, and phosphate were also detected. LPS from bdellovibrios grown intraperiplasmically contained components characteristic of both axenically grown bdellovibrios and the substrate cells. Substrate cell-derived LPS fatty acids made up the majority of the bdellovibrio LPS fatty acids and were present in about the same proportions as in the substrate cell LPS. Glucosamine derived from E. coli LPS amounted to about one-third of the hexosamine residues in intraperiplasmically grown bdellovibrio LPS. However, galactose, characteristic of the E. coli outer core and O antigen, was not detected in the bdellovibrio LPS, suggesting that only lipid A components of the substrate cell were incorporated. Substrate cell-derived and bdellovibrio-synthesized LPS materials were conserved in the B. bacteriovorus outer membrane for at least two cycles of intraperiplasmic growth. When bdellovibrios were grown on two different substrate cells successively, lipid A components were taken up from the second while the components incorporated from the lipid A of the first were conserved in the bdellovibrio LPS. The data show that substrate cell lipid A components were incorporated into B. bacteriovorus lipid A during intraperiplasmic growth with little or no change, and that these components, fatty acids and hexosamines, comprised a substantial portion of bdellovibrio lipid A.

Bdellovibrio↗

Partial characterization of lipid A of intraperiplasmically grown Bdellovibrio bacteriovorus.

The lipid A components of substrate cell origin incorporated by Bdellovibrio bacteriovorus during intraperiplasmic growth (D. R. Nelson and S. C. Rittenberg, J. Bacteriol. 147:860-868, 1981) were shown to be integrated into its lipopolysaccharide structure. Lipid A isolated from bdellovibrios grown on Escherichia coli was resolved into two fractions by thin-layer chromatography. Fraction 2 had the same Rf as the single lipid A fraction of axenicaly grown bdellovibrios, and both stained identically with aniline-diphenylamine reagent. Fraction 1 resembled, in Rf and staining reaction, the slower migrating of two lipid A fractions obtained from the E.coli used as the substrate cell. Both fractions 1 and 2 contained glucosamine, a substrate cell-derived compound. Greater than 65% of the fatty acids in fraction 1 were derived from the substrate cell, whereas more than 60% of the fatty acids of fraction 2 were synthesized by the bdellovibrio. Nevertheless, each fraction contained significant amounts of fatty acid of both origins. The substrate cell-derived fatty acids had the same distribution of N-acyl and O-acyl linkages as in E. coli lipid A. The data indicate that the two lipid A moieties in lipopolysaccharide of intraperiplasmically grown bdellovibrios are hybrids of substrate cell-derived and bdellovibrio-synthesized components. The data also suggest that disaccharide units and N- and O-acyl linkages preexisting in the substrate cell lipid A may be conserved. A possible explanation for the unequal distribution of substrate cell-derived material in the two lipid A fractions of the bdellovibrio is suggested.

Bdellovibrio↗

Characterization and distribution of the hydrocarbons found in diapausing pupae tissues of the tobacco hornworm, Manduca sexta (L.).

The cuticular hydrocarbons of diapausing pupae of the tobacco hornworm, Manduca sexta (L.), are composed of two types of hydrocarbons: n-alkanes (3%) and unsaturated hydrocarbons (97%). The integument, fat body, muscle, and hemolymph are composed of three types of hydrocarbons: n-alkanes (9, 6, 9, and 2%, respectively), branched alkanes (75, 85, 80, and 56%, respectively), and unsaturated hydrocarbons (16, 9, 11, and 42%, respectively). The absence of branched alkanes on the cuticular surface indicated that hydrocarbons are selectively synthesized or deposited according to the presence or absence of methyl branches in the molecule. The hydrocarbons consisted of homologous series of n-alkanes from 21 to 41 carbon atoms, monomethylalkanes from 23 to 39 carbon atoms, dimethylalkanes from 25 to 43 carbon atoms, and trimethylalkanes from 33 to 37 carbon atoms. All branched components had methyl groups near the center of the molecule, and the di- and trimethyl branched components had three methylene units between the branch points. Straight chain alkenes, alkadienes, and alkatrienes from 23 to 44 carbon atoms were partially characterized. The percentage composition of each homologous series varied from tissue to tissue.

Alkanes↗

Fertility of goats following synchronization of estrus with prostagland in F2alpha.

Thirty-four mixed breed cyclic does were randomly divided into two groups of 17 each. One group was synchronized for estrus using two i.m. injections of 8 mg PGF2alpha administered 11 days apart. The other group served as controls and was bred at the time of naturally occurring estrus. Both groups were bred by natural service. Ninety-four percent of the treated does came into estrus within a mean (+/- S.E.) of 53 +/- 3 hours after the second injection of 8 mg PGF2alpha. No differences (P > 0.10) in the first service conception rates based on radiography at mid-gestation were observed between the treated and control groups. It was concluded that the use of 8 mg injections of PGF2alpha 11 days apart had no detrimental effects on fertility of goats.

Journal Article↗

Effect of presence of the male on initiation of estrous cycle activity of goats.

Serum progesterone concentrations and behavioral estrus were determined in two groups of 17 mixed breed dairy does at the beginning of the breeding season. The treatment group was pastured adjacent to two mature bucks while two teaser bucks ran with the group. The control group was pastured without exposure to bucks. Goats were observed for estrus daily for 35 days and samples of jugular blood were collected every other day for radioimmunoassay of progesterone. Signs of estrus were observed in 16 of 17 does in the treatment group within a mean +/- S.E. of 5.5 +/- 1.3 days after introduction of the bucks. Thirteen does demonstrated a progesterone profile characteristic of a normal estrous cycle with peak progesterone concentrations of 5.9 +/- 0.5 ng/ml. Signs of behavioral estrus were not observed in the control group. One control doe demonstrated a progesterone profile characteristic of a normal estrous cycle attaining a peak progesterone concentration of 3.9 ng/ml. Progesterone concentrations in the remaining 16 control does were at or near the lower limits of sensitivity of the assay for the duration of the experiment. Fifteen of the control does exhibited estrus within 7 +/- 1.5 days after exposure to bucks at the end of the experiment. These results clearly demonstrated a profound influence of the male on estrous cycle activity during the beginning of the breeding season.

Journal Article↗