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Biomedical subjects

K L Anderson

Publications and source records attributed to K L Anderson.

At least 109 records · Page 6Linked to original sources

A naturally occurring horizontal gene transfer from a eukaryote to a prokaryote.

Naturally occurring horizontal gene transfers between nonviral organisms are difficult to prove. Only with the availability of sequence data from a wide variety of organisms can a convincing case be made. In the case of putative gene transfers between prokaryotes and eukaryotes, the minimum requirements for inferring such an event include (1) sequences of the transferred gene or its product from several appropriately divergent eukaryotes and several prokaryotes, and (2) a similar set of sequences from the same (or closely related organisms) for another gene or genes. Given these criteria, we believe that a strong case can be made for Escherichia coli having acquired a second glyceraldehyde-3-phosphate dehydrogenase gene from some eukaryotic host. Ancillary observations on the general rate of change and the time of the prokaryote-eukaryote divergence support the notion.

Amino Acid Sequence↗

Pharmacokinetics of flunixin meglumine in lactating cattle after single and multiple intramuscular and intravenous administrations.

The pharmacokinetics of flunixin were studied in 6 adult lactating cattle after administration of single IV and IM doses at 1.1 mg/kg of body weight. A crossover design was used, with route of first administration in each cow determined randomly. Plasma and milk concentrations of total flunixin were determined by use of high-pressure liquid chromatography, using an assay with a lower limit of detection of 50 ng of flunixin/ml. The pharmacokinetics of flunixin were best described by a 2-compartment, open model. After IV administration, mean plasma flunixin concentrations rapidly decreased from initial concentrations of greater than 10 micrograms/ml to nondetectable concentrations at 12 hours after administration. The distribution phase was short (t1/2 alpha, harmonic mean = 0.16 hours) and the elimination phase was more prolonged (t1/2 beta, harmonic mean = 3.14 hours). Mean +/- SD clearance after IV administration was 2.51 +/- 0.96 ml/kg/min. After IM administration, the harmonic mean for the elimination phase (t1/2 beta) was prolonged at 5.20 hours. Bioavailability after IM dosing gave a mean +/- SD (n = 5) of 76.0 +/- 28.0%. Adult, lactating cows (n = 6) were challenge inoculated with endotoxin as a model of acute coliform mastitis. After multiple administration (total of 7 doses; first IV, remainder IM) of 1.1 mg/kg doses of flunixin at 8-hour intervals, plasma flunixin concentrations were approximately 1 microgram/ml at 2 hours after each dosing and 0.5 micrograms/ml just prior to each dosing. Flunixin was not detected in milk at any sampling during the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Analysis of human T cell response to two Plasmodium falciparum merozoite surface antigens.

Eight novel human T cell epitopes were identified within the two major merozoite surface antigens (MSA1 and MSA2) of Plasmodium falciparum using synthetic peptides. All except one of the peptides conformed structurally to an amphipathic alpha helix and three out of the four MSA1 peptides also contained sequences containing the Rothbard motif. Peptide MSA2/2, which fitted none of these criteria, was recognized by our donors to a similar degree as the other peptides. This peptide also contains a B cell epitope. Proliferative responses were obtained in both immune and nonimmune donors, however, the number of responses in the immune donor group was significantly higher. There was no correlation between the level of proliferation and antibody titers to these antigens. No peptides were preferentially recognized in association with specific HLA class II antigens.

Animals↗

Anti-inflammatory therapy in acute endotoxin-induced bovine mastitis.

Acute mastitis was induced in lactating cows by intramammary challenge with 10 micrograms of Escherichia coli lipopolysaccharide. The cows were monitored clinically prior to and for 96 hours after challenge. Milk production, complete blood counts, serum enzyme activities and milk indicators of inflammation were evaluated. Endotoxin challenge was in 7 groups of 3 cows each. Within the groups, cows were randomly assigned to 3 intravenous treatments: saline controls, steroid (one dose of dexamethasone at 0.44 mg/kg) and non-steroidal agent (two doses of flunixin meglumine at 1.1 mg/kg, 8 h apart). Anti-inflammatory therapy reduced rectal and mammary gland surface temperatures. Milk production was significantly reduced (p less than 0.05) in cows treated with dexamethasone. Although dexamethasone treatment produced significant increases (p less than 0.05) in blood leukocytes and segmented neutrophils, milk somatic cell concentrations were not significantly altered. Flunixin meglumine did not alter milk production or blood or milk leukocytes.

Acute Disease↗

Biochemical evidence that starch breakdown by Bacteroides thetaiotaomicron involves outer membrane starch-binding sites and periplasmic starch-degrading enzymes.

Bacteroides thetaiotaomicron can utilize amylose, amylopectin, and pullulan as sole sources of carbon and energy. The enzymes that degrade these polysaccharides were found to be primarily cell associated rather than extracellular. Although some activity was detected in extracellular fluid, this appeared to be the result of cell lysis. The cell-associated amylase, amylopectinase, and pullulanase activities partitioned similarly to the periplasmic marker, acid phosphatase, when cells were exposed to a cold-shock treatment. Two other enzymes associated with starch breakdown, alpha-glucosidase and maltase, appeared to be located in the cytoplasm. Intact cells of B. thetaiotaomicron were found to bind 14C-starch. Binding was probably mediated by a protein because it was saturable and was decreased by treatment of cells with proteinase K. Results of competition experiments showed that the starch-binding proteins had a preference for maltodextrins larger than maltohexaose and a low affinity for maltose and maltotriose. Both the degradative enzymes and starch binding were induced by maltose. These findings indicate that starch utilization by B. thetaiotaomicron apparently does not involve secretion of extracellular enzymes. Rather, binding of the starch molecule to the cell surface appears to be a first step to passing the molecule through the outer membrane and into the periplasmic space.

Bacterial Outer Membrane Proteins↗

Genetic evidence that outer membrane binding of starch is required for starch utilization by Bacteroides thetaiotaomicron.

Mutagenesis of Bacteroides thetaiotaomicron with the transposon Tn4351 produced five classes of mutants that were not able to grow on amylose or amylopectin. These classes of mutants differed in their ability to grow on maltoheptaose (G7) and in the level of starch-degrading enzymes produced when bacteria were grown on maltose. All of the mutants were deficient in starch binding. Since one class of mutants retained normal levels of starch-degrading enzymes, this indicates that binding of the starch molecule by a cell surface receptor is necessary for starch utilization by B. thetaiotaomicron. Analysis of a starch-negative mutant that grew on G7 indicated that B. thetaiotaomicron possessed two starch-binding components or sites. One component (site A), apparently missing in this mutant, had an absolute preference for larger starch oligomers, whereas the other component (site M) also had a high affinity for maltodextrins (G4 through G7). Mutants not able to grow on maltodextrins (greater than G4) probably lacked both of these binding components. Only one class of mutants did not grow normally on maltose, but instead had a 4- to 5-h lag on maltose and a slower growth rate than the wild type. This class of mutants did not produce any of the starch-degrading enzymes or bind starch, even when growing on maltose. Such a phenotype probably resulted from transposon inactivation of a central regulatory gene or a gene encoding an enzyme that produces the inducer. The fact that both the degradative enzymes and the starch-binding activity were affected in this mutant indicates that genes encoding the cell surface starch-binding site are under the same regulatory control as genes encoding the enzymes.

Bacterial Outer Membrane Proteins↗

Identification of T epitopes within a potential Plasmodium falciparum vaccine antigen. A study of human lymphocyte responses to repeat and nonrepeat regions of Pf155/RESA.

PBMC from Melanesians who had high antibody reactivities to fusion proteins encompassing the 3' and the 5' repeat regions of the ring infected E surface antigen (Pf155/RESA), were tested for their ability to respond to synthetic and recombinant peptides representing regions of Pf155/RESA. The aim was to identify T cell epitopes within the Ag. Most of the synthetic peptides from the nonrepeat regions of Pf155/RESA were selected for study on the basis of their tendency to form amphipathic alpha-helices. Peptides representing immunodominant B cell epitopes were also tested. Three-quarters of the Melanesian donors responded to the recombinant peptides (Ag 1505 and Ag 632-100) and to the 8 x 4 mer, a synthetic peptide representative of the 3' repeat region. Whereas all the remaining eight peptides tested elicited a response in at least one donor, three peptides (M40, M42, and BTA3) representing sequences in the nonrepeat regions showed greatest promise as potentially useful T epitopes. Responses in control donors were also observed to most of the peptides but the percentage of responders was lower. T cell bulk lines specific to Ag 1505 and Ag 632-100 were established. All donors were HLA tissue typed, but no obvious correlations between responsiveness and HLA type were observed. Our results suggest that there are T cell epitopes within and outside the repeat regions of Pf155/RESA.

Adult↗

Sources of Prototheca spp in a dairy herd environment.

A source of Prototheca spp causing mastitis in a herd of 263 milking cows in North Carolina was investigated. Of 38 samples from the dairy environment, 18 (47%) contained Prototheca spp. Isolation sites included cattle drinking water; a feed trough; mud, dirt, and excreted feces from a dirt lounging area; water, sludge, mud, and vegetation from a creek in the lounging area; and the floor of a freestall barn. Samples were collected from 5 additional dairies, including one dairy with and 4 dairies without a history of protothecal mastitis. Prototheca spp were isolated from 48 (25%) of 190 samples from various sites on all 6 dairies. Isolates were P zopfii (45; 94%) and P wickerhamii (3; 6%). Isolation frequency ranged from 4 to 47% of samples/dairy. There was no apparent difference in the isolation frequency of Prototheca spp from samples from dairies with or without a history of protothecal mastitis. Sites characterized by wetness and the presence of organic matter most commonly yielded Prototheca spp. Because Prototheca spp appear to be common in the dairy environment, factors in addition to presence in the environment may be important in development of protothecal mastitis.

Animals↗

Immunostimulatory capabilities of highly enriched Langerhans cells in vitro.

Langerhans cells (LC) are epidermal antigen-presenting cells capable of inducing allogenic, antigen-specific, and cytotoxic T cell proliferation. Previous studies have examined the dynamics of LC maintained in vitro in crude epidermal cell (EC) suspensions in which the major cell type is the keratinocyte (KC). To avoid the confounding effects of KC and other immunoregulatory cells on LC dynamics in vitro, highly enriched murine LC (85%) were studied, through 72 h of incubation in vitro, for their ability to present alloantigen (in a primary allogenic proliferation assay) and foreign antigen (in a secondary autologous proliferation assay). The results were compared to similar studies using crude EC suspensions. Freshly prepared LC are very poor stimulators of a primary allogenic proliferation response, with a 12- to 16-fold increase in stimulatory capacity by 72 h using panned-enriched and crude EC suspensions, respectively. Similarly, freshly prepared LC are weak stimulators of a secondary autologous proliferation response, with a 2.5- to 6-fold increase in immunostimulatory capability by 72 h. The overall increased stimulatory effect observed with the crude EC suspensions compared to highly enriched LC is most likely attributed to the effect of KC on T cell proliferation, rather than to a maturation effect of KC on LC during the 72 h of in vitro incubation. Using back-scattered electron imaging, the surface density of MHC-class II molecules (Ia) increased three- to fourfold through culture, which parallels the increase in functional ability. This study demonstrates that LC in either a crude or highly enriched cell suspension mature into potent immunostimulatory cells after incubation in vitro with an increased surface expression of Ia molecules. Keratinocytes are not necessary for LC maturation in vitro, but seem to exert some stimulatory effect by enhancing lymphocyte proliferation in the functional assay system.

Animals↗

Performance and ruminal changes of early-weaned calves fed lasalocid.

Twenty-two neonatal calves were assigned to a control or lasalocid-fed group and weaned at 3 wk of age. They were fed a prestarter diet from 3 d of age until they consumed 227 g/d and then a mixture of 227 g prestarter daily and starter diet in ad libitum amounts. The lasalocid-fed group received lasalocid in milk at 1 mg/kg body weight daily from 4 to 7 d and at .5 mg/kg body weight daily in milk and medicated prestarter diet (88 mg lasalocid/kg) during the 2nd wk. After 2 wk, lasalocid-fed calves were given medicated prestarter and starter (44 mg lasalocid/kg) diets. Four calves in each group were ruminally cannulated at 3 to 5 d of age, and ruminal contents were obtained at weekly intervals to monitor microbial activity. Rectal fecal samples were collected from all calves and examined for coccidial oocysts. Lasalocid-fed calves had a greater weekly feed intake and weight gain than control calves after 6 wk of age. Total ruminal volatile fatty acid concentrations were higher, but the acetate:propionate ratio was lower in lasalocid-fed calves than in control calves. Total viable anaerobic and amylolytic bacterial counts were higher in lasalocid-fed calves than in control calves. No significant treatment effect was found for ruminal NH3-N concentration or ruminal lasalocid-resistant, lactobacilli, lactate-utilizing, cellulolytic or methanogenic bacterial numbers. No evidence of coccidiosis was detected in either group. In general, lasalocid-fed calves had greater feed intake, weight gain and ruminal microbial activity than the calves fed no lasalocid in the diet.

Animals↗

Formation and fate of fermentation products in hot spring cyanobacterial mats.

The fate of representative fermentation products (acetate, propionate, butyrate, lactate, and ethanol) in hot spring cyanobacterial mats was investigated. The major fate during incubations in the light was photoassimilation by filamentous bacteria resembling Chloroflexus aurantiacus. Some metabolism of all compounds occurred under dark aerobic conditions. Under dark anaerobic conditions, only lactate was oxidized extensively to carbon dioxide. Extended preincubation under dark anaerobic conditions did not enhance anaerobic catabolism of acetate, propionate, or ethanol. Acetogenesis of butyrate was suggested by the hydrogen sensitivity of butyrate conversion to acetate and by the enrichment of butyrate-degrading acetogenic bacteria. Accumulation of fermentation products which were not catabolized under dark anaerobic conditions revealed their importance. Acetate and propionate were the major fermentation products which accumulated in samples collected at temperatures ranging from 50 to 70 degrees C. Other organic acids and alcohols accumulated to a much lesser extent. Fermentation occurred mainly in the top 4 mm of the mat. Exposure to light decreased the accumulation of acetate and presumably of other fermentation products. The importance of interspecies hydrogen transfer was investigated by comparing fermentation product accumulation at a 65 degrees C site, with naturally high hydrogen levels, and a 55 degrees C site, where active methanogenesis prevented significant hydrogen accumulation. There was a greater relative accumulation of reduced products, notably ethanol, in the 65 degrees C mat.

Journal Article↗

Ruminal microbial development in conventionally or early-weaned calves.

Eight bull calves (four Holsteins and four Ayrshire calves with mean birth weight of 38.8 kg) were ruminally cannulated at 3 d of age and allocated to one of two weaning programs. All calves were fed colostrum for 3 d after birth and milk until weaning. Calves in the conventional-weaning program were fed a starter diet from 3 d of age and weaned at 6 wk of age. In the early-weaning program, calves were fed a highly palatable pre-starter diet from 3 d of age until they consumed 227 g/d, and then fed a mixture of pre-starter (227 g) and starter diet ad libitum. Calves in this group were weaned at 4 wk of age. Ruminal samples were collected at 3 and 7 d, then weekly thereafter through 8 wk and at 10 and 12 wk of age to assess microbial activity. Calves in the early-weaned group had a higher concentration of total ruminal volatile fatty acids at an earlier age than the calves in the conventional-weaning program. This was accompanied by a trend toward higher lactate concentrations and lower ruminal pH in the early-weaned group during their first 4 wk of age. Lactate and ammonia concentrations decreased with calf age. The total anaerobic bacterial counts increased slightly with calf age, whereas Streptococcus bovis and facultative bacterial populations decreased with calf age. Amylolytic, proteolytic, lactobacilli, lactate-utilizers, cellulolytic and methanogenic bacterial populations increased progressively in both groups. Cellulolytic and methanogenic bacteria were present in both groups at 3 d of age. No protozoa were detected in calves of either group. In general, the most significant changes in bacterial populations and metabolic activity in both groups occurred between 4 and 6 wk of age. Although calves in both groups had similar patterns of bacterial development, calves in the early-weaning program tended to have high ruminal microbial activity at an earlier age than the conventionally weaned calves.

Animals↗

Ruminal metabolic development in calves weaned conventionally or early.

Eight neonatal bull calves were rumen fistulated and assigned to one of two weaning programs to study the effect of diet and weaning age on ruminal metabolic development. All calves were fed colostrum until 3 d of age and milk until weaning. Calves in the early weaning program were fed milk and a highly palatable, prestarter diet until they consumed 227 g/d and then a mixture of 227 g of prestarter and all the starter diet they would eat. Calves in this group were weaned at 4 wk of age. Calves in the conventional weaning program were fed milk and a starter diet and weaned at 6 wk of age. Eight ruminal samples were collected over 12 h from each calf at 1, 4, 8, and 12 wk of age. Ruminal fluid samples were analyzed for pH, VFA, NH3 N, and L(+)-lactate and D(-)-lactate concentrations. Calves weaned early had lower ruminal pH, higher total VFA concentration, and higher molar proportion of butyrate than conventionally weaned calves. Ruminal NH3 N and lactate concentrations were not significantly affected by the weaning program, although lactate tended to be higher in calves weaned early. Ruminal VFA concentration increased and NH3N concentration decreased with increased feed consumption by calves in both groups. Molar proportions of acetate, isobutyrate, and isovalerate decreased, and those of propionate increased with age in both groups. Apparently, ruminal metabolic development, as evidenced by changes in fermentation products, was faster in calves weaned early than in calves weaned conventionally.

Animals↗

Immune-mediated hemolytic anemia induced by penicillin in horses.

Three horses developed severe, immune-mediated hemolytic anemia after treatment with penicillin. The horses had positive direct antiglobulin (Coombs) tests and high titers of IgG antibody that agglutinated penicillin-coated equine red cells. Two of the horses were tested for antibodies to autologous red cell antigens; autoantibodies were not present. Titers of antipenicillin antibody decreased after penicillin was discontinued but IgG antibody was detectable months after recovery. One of the horses was challenged with penicillin; antibody titer increased slightly, but anemia did not develop. Antipenicillin antibody of the IgM class was present in low titer in 23 (77%) of 30 non-anemic horses tested. Apparently, the horse is similar to man in that penicillin-induced anemia is rare but the percentage of individuals with antipenicillin antibody is high.

Anemia, Hemolytic↗

Androgeny, flexibility, and individualism.

Questionnaire data from 195 college students were examined to clarify the relationships among masculinity, femininity, and androgyny and self-perceived flexibility and individualism. Analysis of variance (ANOVA) and hierarchical regression showed no evidence for unique positive "androgyny" effects, while higher masculinity was associated with flexibility and individualism, particularly among women. Contrary to expectations, "androgynous" men were more rigid and conforming, while the cross-typed were most flexible and independent.

Journal Article↗

Endotoxin-induced bovine mastitis: arachidonic acid metabolites in milk and plasma and effect of flunixin meglumine.

Arachidonic acid metabolites (AAM) were measured in milk and plasma during the course of acute endotoxin-induced mastitis in 12 lactating cows. Mastitis was induced by intramammary challenge exposure with 10 micrograms of Escherichia coli (026:B6) endotoxin. Endotoxin was injected into the teat cistern via the teat canal of a single randomly selected rear quarter of each cow. Concentrations of prostaglandin (PG) F2 alpha and thromboxane (Tx) B2 in fat-free unextracted milk and of 15-keto-13,14-dihydro-PGF2 alpha in plasma were measured by radioimmunoassay. Total production of AAM in milk was determined by measuring quarter milk production. The AAM were compared in 6 cows administered flunixin meglumine (1.1 mg/kg of body weight) and in 6 cows administered saline solution. Concentrations of TxB2 in milk were significantly (P less than 0.001) increased during the early course of acute mastitis in endotoxin-treated quarters of cows not administered flunixin meglumine. Peak concentrations of TxB2 in milk occurred at 8 hours after endotoxin inoculation. Flunixin meglumine treatment produced significant (P less than 0.05) reductions in milk TxB2 and plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations. Concentrations of PGF2 alpha in milk and total PGF2 alpha and TxB2 production per quarter per milking were not significantly influenced by endotoxin challenge or by flunixin meglumine treatment.

Animals↗