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Biomedical subjects

J H Harrison

Publications and source records attributed to J H Harrison.

At least 37 records · Page 2Linked to original sources

Particle development in apple juice determined by light scattering and electron microscopy.

Light scattering was used to detect the kinetic development of particles in apple juices produced with and without oxidation and in procyanidin extracts prepared from these two juices. Particle size developed exponentially in both oxidized and unoxidized juices, suggesting an enzymatic origin for the particle forming reactions. The procyanidin extract from oxidized juices produced particles which grew linearly, suggesting diffusion-controlled aggregation or coalescence of particles. Procyanidins from unoxidized juice showed no particle development over at least 60 days of storage. Electron microscopy showed particles similar to those seen previously and some newly described morphologies. The mechanism of haze particle development is discussed in the light of the light scattering and electron microscopic results.

Beverages↗

Estimation of the flow of microbial nitrogen to the duodenum using urinary uric acid or allantoin.

Data were collected from six experiments using duodenally cannulated Holstein dairy cows (88 combinations of cow and period) to evaluate the relationship between urinary purine metabolites and microbial N flow. Experiments evaluated the effects of dietary factors on microbial N production, which included 1) varying concentrations of ruminally degradable protein and nonstructural carbohydrates, 2) supplemental sources of protected amino acids, 3) grass silage treated with fibrolytic enzymes, 4) bacterial inoculation of corn silage, and 5) ruminal starch availability as affected by corn silages of varying maturity. The coefficient of determination for individual experiments that measured the relationship between microbial N flow and allantoin or uric acid excretion in urine ranged from 0.01 to 0.68 and 0.02 to 0.82, respectively. Across all experiments, the coefficients of determination between microbial N flow and allantoin or uric acid excretion in urine were r2 = 0.002 and 0.11, respectively. Removal of data from one experiment improved the overall coefficient of determination between microbial N flow and urinary uric acid to r2 = 0.32. Urinary allantoin excretion across experiments was negatively correlated with microbial N flow, but urinary allantoin excretion within experiments was positively correlated with microbial N flow. Uric acid excretion in urine was positively correlated with microbial N flow across and within experiments, except for one experiment. Our data demonstrate that uric acid excretion in urine can be used to predict microbial N production, except in early lactation, and that urinary allantoin excretion cannot be used to predict microbial N production accurately among cows at different stages of lactation.

Allantoin↗

The effect of ruminal bypass lysine and methionine on milk yield and composition of lactating cows.

Fifty-six multiparous Holstein cows were assigned at 3 wk prepartum to rations based on grass silage with 1) corn distillers grains to provide 86 and 90% of estimated required metabolizable Lys and Met, respectively; 2) a blend of blood meal, fish meal, and meat and bone meal as amino acid (AA) sources to provide 112 and 103% of required metabolizable Lys and Met, respectively; 3) ruminally protected Lys and Met added as a top-dressing to ration 1 to provide 27 g/d of Lys and 8 g/d of Met as available AA at the duodenum postpartum; and 4) ruminally protected AA for 8 wk postpartum as a top-dressing to ration 1 to provide 40 g/d of Lys and 13 g/d of Met as available AA at the duodenum. Cows fed rations 3 and 4 were offered 13.5 g/d of duodenally available Lys and 4 g/d of duodenally available Met for 3 wk prepartum. The total length of the study was 43 wk. Cows fed ration 4 consumed 3 to 4 kg more dry matter than did cows fed the other three rations, and milk yield and the percentage of milk protein and fat were significantly increased during the first 8 wk of lactation. In early lactation, cows fed ration 3 had a greater milk fat percentage but similar dry matter intake, protein percentage, and yield of 4% fat-corrected milk compared with cows fed ration 2. The concentrations of blood serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, triglyceride, and nonesterified fatty acids were lower for cows fed ration 4 during the first 8 wk of lactation than they were for cows fed the other three rations. The mammary arteriovenous difference of whole blood AA indicated that Met along with His and Arg may be the most limiting AA for milk yield.

Amino Acids↗

The occurrence and replication of Escherichia coli in cattle feeds.

Sixty-three of 209 (30.1%) samples of cattle feed that were collected from multiple commercial sources and from farms were found to contain Escherichia coli. However, none of the feed samples examined were culture-positive for E. coli O157. Replication of fecal E. coli, including E. coli O157, was demonstrated in a variety of feeds at temperatures that were similar to those found on farms in summer months. Fresh mixed rations containing corn silage were sampled from 16 dairies. Rations from 12 of these dairies were found to contain E. coli, and the rations from 5 dairies had concentrations of E. coli that were greater than 1000 cfu/g. The ability of experimental mixed rations to support the replication of E. coli was correlated with the concentration of organic acids in the corn silage that was used in the ration. Widespread contamination of cattle feeds with E. coli and the ability of E. coli to replicate in feeds suggest that feeds are a potentially important factor in the ecology of organisms that can be transmitted from feces to mouth, such as E. coli O157.

Acetates↗

Effect of whole cottonseed, gossypol, and ruminally protected lysine supplementation on milk yield and composition.

To determine whether the gossypol content of the diet affected availability of dietary Lys, 4 cows with ruminal and duodenal cannulas and 16 intact cows in early lactation were assigned to diets containing either glanded cottonseed, glanded cottonseed plus 15 g/d of ruminally protected Lys, glandless cottonseed, or glandless cottonseed plus 15 g/d of ruminally protected Lys. The experimental design was a 4 x 4 Latin square design with 3-wk experimental periods. Gossypol concentrations in plasma were lower when cows were fed glandless cottonseed. Lysine supplementation increased the flow and percentage of Lys in duodenal digesta and increased the concentration of Lys in plasma. Neither type of cottonseed nor Lys supplementation affected dry matter intake. Yields of milk and fat-corrected milk from cannulated cows were not affected by diet, but yields from intact cows were increased by Lys supplementation. For both cannulated and intact cows, Lys supplementation increased the percentage of milk protein, and milk protein yield was increased by Lys in intact cows. Dietary gossypol did not affect the response to Lys, indicating that dietary gossypol had little or no effect on the availability of Lys for dairy cows.

Amino Acids↗

Isolation and primary culture of murine alveolar type II cells.

Previous attempts to culture mouse alveolar type II (ATII) cells have been hampered by limited purity and cell recovery. We have now obtained culturable ATII cells from female C57BL/6 mice at a purity of 92% +/- 3 (mean +/- SD; n = 20), with viabilities of 96% +/- 2 and total yields of 5.1 +/- 0.7 X 10(6) cells per mouse. Crude lung cell suspensions were prepared by intratracheal instillation of Dispase and agarose followed by mechanical disaggregation of the lungs. Crude cell suspensions were purified by negative selection using a biotinylated-antibody, streptavidin-coated biomagnetic particle system. Cell purities were determined by Pap staining and confirmed ultrastructurally. Purified ATII cells were cultured on fibronectin-coated chamber slides and maintained for up to 5 days in DMEM with 10% fetal bovine serum. Cultures exhibited minimal contamination by Clara cells, mesenchymal cells, or endothelial cells, and the epithelial nature of the cultures was confirmed by positive cytokeratin staining in at least 97% of the cells through day 5. Day 3 cultures demonstrated osmium tetroxide/tannic acid-stained granules consistent with lamellar bodies in 76% +/- 3.6 of the cells. The cultures displayed features distinct from those previously described for adult rat ATII cells, including irregularly-shaped cells and the formation of numerous cytoplasmic projections in direct contact with other cells. These studies indicate that excellent yields of highly purified, culturable ATII cells can be obtained from genetically defined mice. These techniques may provide powerful new models for the study of parenchymal lung disease in vitro.

Animals↗

Identification of patients for pharmacologic review by computer analysis of clinical laboratory drug concentration data.

Clinical pharmacokinetics evaluation and consultation can improve drug therapy and decrease the incidence of adverse reactions in selected patients. However, identification of patients appropriate for review is difficult. The authors developed a microcomputer-based expert system that scans clinical laboratory drug concentration data to identify patients for follow-up. Rules were developed from a review of data for digoxin, phenytoin, and theophylline. These were implemented in software that provides for simple rule creation and modification, on-screen graphic review of data, and printing of chart reports. This program is readily adapted for use with most laboratory information systems. In a retrospective study of 868 patients monitored for digoxin, phenytoin, and theophylline, 29% were flagged as having drug level profiles of possible concern. The majority (62%) of these patients had multiple specimens flagged, suggesting persistent problems. These data suggest that patients can be identified for follow-up by scanning serial drug concentrations, allowing consultative resources to be focused on patients most likely to benefit from them.

Clinical Laboratory Information Systems↗

Effect of whole cottonseeds and calcium salts of long-chain fatty acids on performance of lactating dairy cows.

Holstein cows were assigned to diets containing no supplemental fat, supplemental fat from whole cottonseed, or supplemental fat from whole cottonseed plus Ca salts of fatty acids (Megalac). The TMR contained 46% forage and 54% concentrate or mixtures of concentrate and whole cottonseed on a DM basis and were fed from wk 3 through 44 of lactation. The mean fat content of the three diets was 3.0, 4.7, and 6.4% of DM for control, whole cottonseed, and whole cottonseed plus Ca salts of fatty acids, respectively. Supplemental fat increased NEL intake, percentage of milk fat, milk fat production, and rate of recovery of BW and body condition. Supplemental fat decreased milk protein production in early lactation, but not in late lactation. Addition of supplemental fat had no significant effect on ruminal concentration of VFA, NH3 N, or in situ digestibility of fiber. The proportion of unsaturated fatty acids in milk fat was increased with supplemental whole cottonseed or whole cottonseed plus Ca salts of fatty acids. During the first 3 mo of lactation, the proportion increased of fatty acids C14 or less, C16, and C18:2. Proportion of fatty acids C16:1 and C18:1 correspondingly decreased. The change in composition of milk fatty acids during early lactation is consistent with the use of body fat for milk synthesis.

Animal Feed↗

Lipid metabolism in adipose tissue of cows fed high fat diets during lactation.

The adaptations of fat synthesis in adipose tissue to lactational state, rate of milk production, and dietary fat intake were determined for dairy cows. Lipogenesis and esterification were determined in cows of average or high genetic merit for milk production and fed either a control TMR of corn silage, alfalfa, and concentrate (2.5% fat; 1.47 Mcal of NEL/kg); a TMR with whole cottonseeds replacing 12% of the concentrate (4.4% fat; 1.49 Mcal NEL/kg); or a TMR with 12% cottonseeds and 2.7% of Ca salts of fatty acids (6.0% fat; 1.53 Mcal of NEL/kg). Dietary treatments began on d 17 of lactation and continued for 288 d. Lipogenesis and esterification decreased equally from 15 d prepartum to 15 d postpartum in all groups. Cows of high merit had lower rates of lipogenesis and esterification at d 60 than did low merit cows but had higher rates of lipogenesis at d 120. Rates of lipogenesis were decreased by dietary fat treatments. Esterification rates were lowest on the intermediate fat TMR and highest on the highest fat TMR. Lipogenesis was decreased logarithmically by dietary fat intake; this effect was greater as lactation progressed. Adipocyte size and body fat mass decreased during early lactation and then increased for all treatment groups. Supplemental dietary fat reduces de novo synthesis of fatty acid, and this effect increases as lactation progresses.

Adipocytes↗

Effect of buffer addition to high grain total mixed rations on rumen pH, feed intake, milk production, and milk composition.

Lactating Holstein cows (n = 56) were fed high grain TMR in three feeding experiments to determine the effects of two rumen buffers on feed intake, milk production, and milk composition. The basal diet in Experiment 1 was 40% grass silage, 12% whole cottonseed, and 48% concentrate with treatments of 1) control (no added buffer), 2) 1.5% of dietary DM as Alkaten, and 3) 1.5% of dietary DM as Rumen 8. In Experiment 2, diets consisted of 12% corn and sunflower silage, 20% whole cottonseed, and 68% concentrate with treatments as in Experiment 1. The diets in Experiment 3 were identical to those in Experiment 2, except that buffers were increased to 2.2% of dietary DM. The DMI was greater for cows fed buffered diets than control diets in Experiments 2 and 3. Milk production was unaffected by treatment. Milk fat percentage was unaffected by diet in Experiment 1 and averaged 3.8%. Milk fat depression was alleviated for cows fed buffered diets in Experiments 2 and 3. No differences in rumen fluid pH or molar percentage of VFA were significant among treatments.

Animal Feed↗

Fermentation and utilization of grass silage.

The decision to utilize particular forages in support of dairy production should be based on a number of key factors, such as available land base, type of manure management, soil type and topography, climate, and availability of purchased forages and feeds. Because of the complexity and environmental concerns existing in the dairy industry today, decisions about forage and manure management should include whole farm analysis with the aid of computer software. The chemical composition and digestibility of grass are affected more by stage of maturity than by other management factors, such as species, DM, or type of harvest system. The decline in digestibility of nutrients in first growth forage is approximately .55 to .68%/d and is dependent on the method of estimation. The decline in digestible DMI in first growth is .3 to .5%/d. The use of silage additives has become an integral part of forage management, and improvements in DMI and milk production are documented. Particle size and type of harvest equipment significantly affect eating behavior and efficiency of milk production. Wilting of silage results in an increase in DMI and efficiency of microbial protein production.

Animal Feed↗

Effect of somidobove sustained release administration on the lactation performance of dairy cows.

Lactation performance was determined on 190 multiparous Holsteins from five herds supplemented with 0, 320, 640, or 960 mg of somidobove every 28 d. The experiment consisted of 21 d of pretreatment and treatment periods of various lengths, depending upon stage of lactation of animals at first administration. Somidobove beginning in early (28 to 45 d in milk), mid (111 to 166 d in milk), or late (166 to 334 d in milk) stages of lactation consisted of 9, 6, or 3 administrations. Milk and 3.5% FCM yields were increased by each dose of somidobove in all stages. Milk composition and dry matter and energy intakes were similar among treatments within stage. Milk to DMI ratio and milk energy to net energy intake ratio were improved by somidobove. Gain was positive for all treatments, but less in somidobove-supplemented cows. Lower body weight and condition score at the completion of somidobove treatment resulted. For early cows, days to first estrus and days to first breeding were similar; however, total number of inseminations for cows receiving somidobove was twofold greater than control, resulting in a longer calving interval. Results demonstrated efficacy of somidobove administered every 28 d to lactating dairy cattle for increased milk yield.

Animals↗

The use of rapeseed screenings in diets for lactating cows and subsequent effects on milk yield and composition.

This trial was to determine the effect of dietary rapeseed screenings on milk production, milk composition, and feed intake of lactating dairy cows. Pelleted rapeseed screenings were fed at 0, 7, and 14% of diet DM. Assignment of 36 multiparous Holstein cows was based upon parity and previous lactation performance. The experimental period was from parturition through wk 16 of lactation. The control ration had 3.1% ether extract, whereas the ration with 14% rapeseed screenings had 5.4% ether extract. The rapeseed screenings contained 52.2% of C18:1 and 24.2% of C18:2 as percentage of total fatty acids. There were no significant effects of dietary treatment on milk yield or feed intake. There were significant increases in milk fatty acids C18:1 and C18:2 for cows consuming the diets with rapeseed screenings. Treatment did not significantly affect milk protein percentage as measured by Kjeldahl N or by the infrared method. Increased unsaturated fatty acids in the milk of cows fed rapeseed screenings had no apparent effect on infrared fat analysis. There was no interaction of treatment and milk fat analysis method (infrared versus Babcock). Dietary treatment did not affect blood serum nonesterified fatty acid concentrations over the 16-wk period. In conclusion, rapeseed screenings, fed at levels up to 14% of the diet, did not affect milk yield, milk protein, or milk fat percentage but did change milk fatty acid composition.

Amino Acids↗

Stimulation of K-C1 cotransport in rat red cells by a hemolytic anemia-producing metabolite of dapsone.

Dapsone, a sulfone compound used in the treatment of leprosy and, more recently, Pneumocystis carinii pneumonia, produces as a major side effect a hemolytic anemia. This anemia is characterized by oxidation of hemoglobin to methemoglobin and increased splenic uptake of red blood cells. Using a rat model, Grossman and Jollow (J. Pharmacol. Exp. Ther. 244: 118-125, 1988) found that dapsone hydroxylamine (DDS-NOH), a dapsone metabolite, is responsible for its hemolytic effect in vivo. DDS-NOH also promotes hemoglobin binding to SH groups on rat red cell membrane proteins (Budinsky et al., FASEB J. 2: A801, 1988). Since the binding of hemoglobin and other reagents (e.g., N-ethylmaleimide) to membrane SH groups has been associated with increased K transport in red blood cells, we examined the effect of DDS-NOH on K efflux from rat red blood cells in vitro. Cells shrink when exposed to DDS-NOH (100 microM) in media with plasma-like ionic composition. This shrinkage is prevented if extracellular K is raised to 110 mM or if intra- and extracellular Cl are replaced by methylsulfate (MeSO4), suggesting involvement of a K-Cl cotransport pathway. Indeed, 100 microM DDS-NOH produces a 4- to 5-fold increase in K efflux in cells containing Cl but less than a 2-fold increase in cells containing MeSO4. This stimulatory effect is specific for K; Na efflux is slightly inhibited by 100 microM DDS-NOH. The concentrations of DDS-NOH required for half-maximal stimulation of Cl-dependent K efflux (53 microM) is similar to its half-maximal hemolytic concentration in rats (approximately 100 microM). Furthermore, the stimulation of Cl-dependent K efflux by DDS-NOH is greater than 80% reversed by subsequent treatment of the cells with dithiothreitol, suggesting involvement of SH groups. Our results indicate that DDS-NOH exposure stimulates an apparent K-Cl cotransport in rat red blood cells, resulting in cell shrinkage under physiological ionic conditions. Since shrinkage of red blood cells renders them less deformable (Mohandas et al., J. Clin. Invest. 66: 563-573, 1980), this suggests a pathophysiological mechanism whereby DDS-NOH exposure in vivo could promote increased splenic uptake of red blood cells and hemolytic anemia.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Effect of inoculation rate of selected strains of lactic acid bacteria on fermentation and in vitro digestibility of grass-legume forage.

Grass-legume forage was used to evaluate the effect of inoculation rate of selected strains of lactic acid bacteria on fermentation and in vitro digestibility during 57 d of ensiling. Chopped forage (DM = 28%) was ensiled in 4 to 6-kg quantities and treated as: 1) control, 10(3) epiphytic lactic acid bacteria; 2) 10(5) added lactic acid bacteria; and 3) 10(6) added lactic bacteria/g of wet forage. Samples were obtained for analyses on d 0, 1, 2, 3, 6, 10, 14, 29, and 57 of fermentation. Treated silages were observed to have: 1) greater quantities of lactic acid bacteria, 2) a greater proportion of homofermentative lactic acid bacteria, and 3) lactic acid bacteria with greater biological activity. Addition of each amount of lactic acid bacteria: 1) increased the rate of utilization of water-soluble carbohydrate and decline in pH, 2) limited the formation of NH3 N, and 3) increased the in vitro digestibility of DM and ADF. No differences were observed in the lactic acid content of the silages after 57 d of fermentation.

Ammonia↗

Acute pulmonary toxicity of bleomycin: DNA scission and matrix protein mRNA levels in bleomycin-sensitive and -resistant strains of mice.

The severity of bleomycin (BLM)-induced pulmonary fibrosis in mice varies markedly among several different murine strains. We have examined the DNA from lungs of sensitive (i.e., C57BL/6N) and resistant (i.e., BALB/c) strains of mice using a nucleoid sedimentation technique to detect early in vivo changes in the integrity of DNA after intravenous BLM. Mice received intravenous injections of BLM (80 mg/kg) or vehicle; lung nucleoids were prepared 15 min to 6 hr later. BLM produced striking decreases in nucleoid sedimentation distance versus paired controls in both strains within 15 min after injection, indicating extensive DNA scission. Repair of DNA strand breaks was complete in the resistant (BALB/c) mice by 5 hr; in contrast, only partial repair occurred in the sensitive (C57BL/6N) strain during that time. We then examined lungs for subsequent changes in steady state poly-(A)+ RNA levels and mRNA levels for lung matrix proteins (type I procollagen, type III procollagen, and fibronectin). Steady state levels of poly-(A)+ RNA were depressed to 50% of control 1 through 6 days after BLM injection in the lungs of sensitive mice. Resistant mice had pulmonary poly-(A)+ RNA levels similar to those of C57BL/6N mice, except for a 2-fold elevation 1 day after BLM injection. BLM injection affected the steady state levels of mRNA encoding lung matrix proteins differently than total poly-(A)+ RNA. Fibronectin mRNA/poly(A)+ RNA was elevated 2-fold 1 day after BLM treatment only in the sensitive strain and remained elevated at 3 and 6 days. In contrast, alpha 2I procollagen mRNA increased in both murine strains and alpha 1III procollagen mRNA decreased in both strains. Thus, a 7-fold or greater increase in the type I: type III procollagen mRNA ratio was seen in both strains 3 to 6 days after BLM injection. These data demonstrate that BLM treatment rapidly produces extensive pulmonary DNA damage in vivo, that persistence of DNA damage rather than the initial level of strand scission is associated with sensitivity to BLM lung disease in these mice, and that changes in the levels of mRNA encoding pulmonary matrix proteins occur in vivo within 1 to 3 days after intravenous BLM treatment.

Animals↗

The reaction of bovine alpha-thrombin with tetranitromethane. Characterization of the modified protein.

Previous studies from several laboratories have shown that thrombin is inactivated by tetranitromethane with the formation of nitrotyrosine. The inactivation is characterized by an apparently greater loss of fibrinogen-clotting activity than activity toward synthetic ester substrates, suggesting that the residues modified by tetranitromethane are involved in the interaction of thrombin with fibrinogen. This study was designed 1) to determine the effect of solvent conditions on the rate of modification and the stoichiometry of the reaction of tetranitromethane with bovine alpha-thrombin; 2) to identify the residue(s) modified; and 3) to characterize the modified enzyme with respect to its interaction with peptide nitroanilide substrates and fibrinogen. The inactivation of thrombin by tetranitromethane proceeded more rapidly in 50 mM Tris, pH 8.0, than in 50 mM sodium phosphate, 100 mM NaCl, pH 8.0. Approximately 10% fibrinogen-clotting activity remained at maximal inactivation. A study of the effect of tetranitromethane concentration on the rate of inactivation suggested that the loss of activity was the result of the modification of 1 mol of tyrosine/mol of thrombin. A similar result was obtained from the analysis of the extent of inactivation as a function of the extent of protein modification. Structural analysis of the modified protein showed substantial modification at both Tyr71 and Tyr85. Enzyme kinetic studies were performed with the modified protein and a control thrombin with N2-tosylglycylprolylarginine p-nitroanilide. H-D-phenylalanylpipecolylarginine p-nitronailide, and purified bovine fibrinogen. With all three substrates, a substantial decrease in kcat was observed, whereas there was essentially no change in Km. These results suggest that, contrary to previous suggestions, the modification of Tyr71 and Tyr85 in thrombin does not influence the binding of substrates, but rather influences active site reactivity.

Animals↗