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Biomedical subjects

L M Johnson

Publications and source records attributed to L M Johnson.

At least 19 recordsLinked to original sources

Palmitate potentiation of glucose-induced insulin release: a study using 2-bromopalmitate.

The mechanisms whereby fatty acids (FA) potentiate glucose-induced insulin secretion from the pancreatic beta cell are incompletely understood. In this study, the effects of palmitate on insulin secretion were investigated in isolated rat islets. Palmitate did not initiate insulin secretion at nonstimulatory glucose concentrations, but markedly stimulated insulin release at concentrations of glucose > or = 5.6 mmol/L. At concentrations of palmitate > or =0.5 mmol/L, the important determinant of the potency of the FA was its unbound concentration. At total concentrations < or = 0.5 mmol/L, both the total and unbound concentrations appeared important. Surprisingly, 2-bromopalmitate did not affect palmitate oxidation, but significantly diminished palmitate esterification into cellular lipids. Neither methyl palmitate, which is not activated into a long-chain acyl-CoA ester, nor 2-bromopalmitate affected glucose-stimulated insulin release. Further, 2-bromopalmitate partly inhibited the potentiating effect of palmitate. These results support the concept that FA potentiation of insulin release is mediated by FA-derived signals generated in the esterification pathway.

Animals↗

Corn silage management: effects of hybrid, chop length, and mechanical processing on digestion and energy content.

Two experiments were conducted to evaluate the effects of chop length and mechanical processing of two hybrids of whole plant corn on digestion and energy content of the total mixed ration (TMR). The experimental designs in experiments 1 and 2 were 6 x 6 and 4 x 4 Latin squares, respectively. In the first experiment, Pioneer hybrid 3845 was harvested at three theoretical lengths of cut: 11.1, 27.8, and 39.7 mm. At each chop length, corn was harvested with and without mechanical processing using a John Deere 5830 harvester with an onboard kernel processor. In the second experiment, Pioneer hybrid Quanta was harvested at two theoretical lengths of cut: 27.8 and 39.7 mm, with and without mechanical processing. In both experiments, the increase in the theoretical length of cut of corn silage increased the average length of cut and tended to increase the percentage of particles greater than 19 mm and lower the percentage of particles between 8 and 19 mm. In experiment 1, apparent total tract dry matter, organic matter, and neutral detergent fiber (NDF) digestibilities were lower for cows fed diets containing corn silage harvested at a short chop length (11.1 mm) than for corn silage harvested at a long chop length (39.7 mm). The lower total tract digestibility of nutrients may have contributed to the lower TDN, metabolizable energy (percentage of digestible energy), and NEL concentration of diets containing the short chop length corn silage (experiment 1). In experiment 2, total tract starch digestibility was greater for cows fed medium chop (27.8 mm) corn silage diets, and total tract NDF digestibility was greater for cows fed long chop (39.7 mm) corn silage diets. The opposing effect of total tract starch and fiber digestibilities between chop lengths may have contributed to the lack of difference in energy content of the diets in experiment 2. The TDN and NEL concentrations of the processed corn silage diets were greater than the unprocessed corn silage diets in experiment 1. The increase in energy concentration for the processed corn silage diet was due to greater total tract digestibility of organic matter and ether extract. Total tract starch digestibility was greater, and total tract NDF digestibility was lower for cows fed processed corn silage diets than unprocessed corn silage diets in experiment 2. The opposing effect of total tract starch and fiber digestibilities between processed and unprocessed corn silage may have contributed to the lack of difference in energy content of the diets.

Animal Feed↗

Corn silage management: effects of hybrid, maturity, inoculation, and mechanical processing on fermentation characteristics.

Two experiments were conducted to evaluate the effects of hybrid, maturity, mechanical processing, and inoculation of corn silage on fermentation characteristics. In experiment 1, Pioneer hybrid 3845 corn silage was harvested at three maturities (hard dough, one-third milkline, two-thirds milkline). In experiment 2, Pioneer hybrids 3845 and Quanta were harvested at three maturities (one-third milkline, two-thirds milkline, and blackline). In both experiments, corn silage was harvested at each maturity with and without mechanical processing and with and without inoculation. In experiments 1 and 2, corn silage was harvested at a theoretical length-of-cut of 6.4 and 12.7 mm, respectively. Maturity at harvest tended to have a greater impact on silage fermentation characteristics of corn silage than mechanical processing and inoculation. In experiments 1 and 2, corn silage harvested at the earliest maturity tended to have decreased dry matter content and increased water-soluble carbohydrate concentrations during the ensiling process than corn silage harvested at advanced maturities. In experiment 2, pH levels were lower for corn silage harvested at the early maturity (one-third milkline) compared with advanced maturities (two-thirds milkline and blackline) by 57 d after ensiling. The difference in pH can be explained by the greater concentration of water-soluble carbohydrates at the early maturity (one-third ML) soon after ensiling (2, 3, 6 and 10 d after ensiling) compared with advanced maturities (two-thirds ML and BL). The increased water-soluble carbohydrate concentrations in the less mature corn silage provided nutrients for bacteria to grow and produce primarily lactic acid (6, 10, and 57 d after ensiling) and some acetic acid (2, 3, 6, and 10 d after ensiling) which reduced the pH of corn silage more than at the advanced maturities. There was a slight change in silage fermentation characteristics when corn silage was inoculated with Pioneer 1132 inoculant in experiment 1. The inoculated corn silage had increased temperature, lactate and acetate concentrations, and lower water-soluble carbohydrate and pH levels compared with uninoculated corn silage. Dry matter recovery tended to be greater for processed corn silage in experiment 1, and greater for unprocessed corn silage in experiment 2. It appears that when fermentation was greater (increased temperature and lactate concentration 57 d after ensiling) the dry matter recovery was lower.

Acetic Acid↗

Corn silage management: effects of hybrid, maturity, chop length, and mechanical processing on rate and extent of digestion.

Five experiments were conducted to evaluate the effects of hybrid, chop length, maturity, and mechanical processing of corn silage on dry matter and nutrient disappearance in the rumen. Corn silage that had not been dried or ground was incubated in macro in situ bags (30 x 35 cm) for 8, 16, 24,48, and 96 h. Experiments la and 2 evaluated the effects of maturity and mechanical processing for two corn silage hybrids harvested at two theoretical lengths-of-cut. Experiments 3 through 5 evaluated the effects of chop length and mechanical processing for two corn silage hybrids harvested at two-thirds milkline. The hybrid with low neutral fiber (NDF) concentrations had greater dry matter and starch disappearance than the hybrid with high NDF concentrations. The effect of chop length on nutrient disappearance was variable across experiments 3 through 5. Processing improved dry matter disappearance in experiments la, 2 (two-thirds milkline and blackline), 3, 4, and 5 at the majority of ruminal incubation timepoints. Starch disappearance was greater for unprocessed corn silage in experiment la (hard dough and two-thirds milkline) and was greater for processed corn silage in experiments 2 and 5. However, there was no consistent trend in starch disappearance between processed and unprocessed corn silage in experiments 3 and 4. This can be partially explained by the high disappearance of starch (experiment 3, > or = 98% and experiment 4, > or = 94%) by 24 h of ruminal incubation. Minimal differences were detected in NDF disappearance between processed and unprocessed corn silage across maturities. In experiments 2 and 5, crude protein disappearance was improved due to processing at some incubation timepoints. Rate of dry matter, starch (one-third milkline and two-thirds milkline), and NDF disappearance tended to increase when corn silage was mechanically processed in experiment 2. Dry matter, starch, and crude protein disappearance tended to be greater for corn silage harvested at the early maturity (one-third milkline) in experiment 2 compared with advanced maturities (two-thirds milkline and blackline). Disappearance of NDF was greater at early maturities compared with advanced maturities in experiments 1a and 2. Rate of dry matter (hybrid 3845), starch, and NDF (hybrid 3845) disappearance tended to decrease as maturity advanced from one-third milkline to blackline in experiment 2. These results suggest that the macro in situ method can be used to evaluate nutrient disappearance in the rumen.

Animals↗

Dose related association of impact activity and bone mineral density in pre-pubertal girls.

Impact activity is an important contributor to bone accrual. Children engaging in such activity have been shown to have greater bone mineral density (BMD) than their peers. This cross-sectional study was designed to quantify the association between BMD and impact activity in pre-pubescent girls, specifically examining for a dose-dependent relationship. Fifty 7-11-year-old Caucasian female gymnasts were grouped by hours of gymnastics participation during a 6-month period: LOW, 1-8 hours/week (hrs/wk); HIGH, >8 hrs/wk. They were compared with 20 controls, with height, weight, age, and Tanner stage averages matched to the gymnasts. Total body, forearm, hip, lumbar spine BMD, and body composition were measured by dual energy X-ray absorptiometry (DXA). Strength was measured by one repetition maximum testing, calcium intake was quantified by questionnaire, and physical activity was measured by questionnaire and activity monitors. Total and regional BMD were greater in the HIGH group than the controls (P<0.05). Furthermore, both total and forearm BMD were greater in the HIGH group than in the LOW group, and greater in the LOW group than in the controls (P<0.05). Simple regression analysis between hrs/wk of gymnastics activity versus total and regional BMD (using maturation-adjusted Z scores) yielded a positive slope for each site. R2 was greatest for hip BMD (R2 = 0.25), and least for lumbar spine (R2 = 0.10) (P<0.0001). In conclusion, in this group of pre-pubescent girls, we observed a dose-dependent relationship between BMD and hrs/wk of impact activity; even moderate doses of impact activity were associated with increased BMD.

Bone Density↗

Implication of alpha4 phosphoprotein and the rapamycin-sensitive mammalian target-of-rapamycin pathway in prolactin receptor signalling.

A prolactin (PRL)-responsive 3'-end cDNA encoding rat alpha4 phosphoprotein was previously isolated from a rat lymphoma cDNA library. Rat alpha4 is a homologue of yeast Tap42 and is a component of the mammalian target-of-rapamycin (mTOR) signalling pathway that stimulates translation initiation and G1 progression in response to nutrients and growth factors. In the present study, the full-length rat alpha4 cDNA was obtained by 5'-RACE and the 1023 bp open reading frame predicted a 340 amino acid protein of 39.1 kDa. The alpha4 mRNA was expressed in quiescent PRL-dependent Nb2 lymphoma cells deprived of PRL for up to 72 h but expression was downregulated within 4 h of PRL treatment. In contrast, PRL-independent Nb2-Sp cells showed constitutive expression of alpha4 that was not affected by PRL. Western analysis of Nb2 cell lysates or of V5-tagged-alpha4 expressed in COS-1 cells detected a single immunoreactive band of approximately 45 kDa. Enzymatic deglycosylation of affinity-purified 45 kDa alpha4 yielded the predicted 39 kDa protein. Phosphorylation of Nb2 alpha4 was induced by PRL or 2-O-tetradecanoyl-phorbol-13-acetate (TPA) and further enhanced by a combination of PRL and TPA. The Nb2 alpha4 associated with the catalytic subunit of protein phosphatase 2A and localized predominantly in Nb2 nuclear fractions with trace amounts in the cytosol. The immunosuppressant drug rapamycin inhibited proliferation of Nb2 cells in response to PRL or interleukin-2, but had no effect on Nb2-Sp cells. Furthermore, transient overexpression of alpha4 in COS-1 cells inhibited PRL stimulation of the immediate-early gene interferon regulatory factor-1 promoter activity. Therefore, PRL downregulation of alpha4 expression and/or PRL-inducible phosphorylation of alpha4 may be necessary for PRL receptor (PRLr) signalling to the interferon regulatory factor-1 promoter in the Nb2 cells and, furthermore, implicates cross-talk between the mTOR and PRLr signalling cascades during Nb2 cell mitogenesis.

Adaptor Proteins, Signal Transducing↗

Corn silage management: effects of maturity, inoculation, and mechanical processing on pack density and aerobic stability.

Three experiments were conducted to evaluate the effects of inoculation, maturity, and mechanical processing of corn silage on aerobic stability and pack density. Corn silage was stored in 20-L mini silos for the three aerobic stability experiments. Corn silage was stored in 80-L mini silos for the three pack-density experiments. The wet pack density of corn silage tended to decrease as maturity advanced in all of the pack-density experiments, and processed corn silage had a greater wet pack density compared with unprocessed corn silage in two of the three 20-L mini silo experiments. Aerobic stability, measured as the number of hours to reach 1.7 degrees C above ambient, was greater for processed corn silage in two of the three 20-L mini silo experiments, and was greater for inoculated corn silage across the three 20-L mini silo experiments. Inoculation of corn silage with lactic acid producing bacteria tended to improve aerobic stability of corn silage more than maturity and mechanical processing.

Aerobiosis↗

Corn silage management I: effects of hybrid, maturity, and mechanical processing on chemical and physical characteristics.

Two experiments were conducted to evaluate the effects of hybrid, maturity, and mechanical processing of whole plant corn on chemical and physical characteristics, particle size, pack density, and dry matter recovery. In the first experiment, hybrid 3845 whole plant corn was harvested at hard dough, one-third milkline, and two-thirds milkline with a theoretical length-of-cut of 6.4 mm. In the second experiment, hybrids 3845 and Quanta were harvested at one-third milkline, two-thirds milkline, and blackline stages of maturity with a theoretical length-of-cut of 12.7 mm. At each stage of maturity, corn was harvested with and without mechanical processing by using a John Deere 5830 harvester with an onboard kernel processor. The percentage of intact corn kernels present in unprocessed corn silage explained 62% of variation in total tract starch digestibility. As the amount of intact kernels increased, total tract starch digestibility decreased. Post-ensiled vitreousness of corn kernels within the corn silage explained 31 and 48% of the variation of total tract starch digestibility for processed and unprocessed treatments, respectively. For a given amount of vitreous starch in corn kernels, total tract starch digestibility was lower for cows fed unprocessed corn silage compared with processed corn silage. This suggests that processing corn silage disrupts the dense protein matrix within the corn kernel where starch is embedded, therefore making the starch more available for digestion. Particle size of corn silage and orts that contained corn silage was reduced when it was processed. Wet pack density was greater for processed compared with unprocessed corn silage.

Animal Feed↗

Corn silage management II: effects of hybrid, maturity, and mechanical processing on digestion and energy content.

Two experiments were conducted to evaluate the effects of maturity and mechanical processing of two hybrids of whole plant corn on starch, fiber, and ether extract digestibilities and energy content of the total mixed ration fed to lactating Holstein cows. In the first experiment, Pioneer hybrid 3845 whole plant corn was harvested at hard dough, one-third milkline, and two-thirds milkline with a theoretical length of cut of 6.4 mm. At each stage of maturity, corn was harvested with and without mechanical processing. In the second experiment, Pioneer hybrids 3845 and Quanta were harvested at one-third milkline, two-thirds milkline, and blackline stages of maturity with and without mechanical processing. The theoretical length of cut was 12.7 mm. The measured TDN and NEL concentrations were lower for diets containing processed corn silage in experiment 1 and greater for diets containing processed corn silage in experiment 2, compared with diets containing unprocessed corn silage. The lower energy content for diets containing processed corn silage in experiment 1 can be explained by the lower total tract NDF, ether extract, and CP digestibilities. The greater energy content for diets containing processed corn silage in experiment 2 can be attributed to greater total tract starch and NDF digestibilities for cows fed processed corn silage diets. In experiment 2, diets containing processed corn silage (1.59 Mcal/kg) had approximately 2.6% more energy available per kilogram of DM consumed compared with diets containing unprocessed corn silage (1.55 Mcal/kg). For hybrid Quanta in experiment 2, the TDN and NEL concentrations of diets containing corn silage harvested at two-thirds ML were greater than at other maturities.

Animal Nutritional Physiological Phenomena↗

Corn silage management III: effects of hybrid, maturity, and processing on nitrogen metabolism and ruminal fermentation.

Two experiments were conducted to evaluate the effects of maturity and mechanical processing of two hybrids of whole plant corn silage on DM and OM digestibility, nitrogen metabolism, ruminal fermentation, and milk production and composition in lactating Holstein cows. In the first experiment, Pioneer hybrid 3845 whole plant corn was harvested at hard dough, one-third milkline, and two-thirds milkline with a theoretical length-of-cut of 6.4 mm. At each stage of maturity, corn was harvested with (1-mm roll clearance) and without (15.9-mm roll clearance) mechanical processing using a John Deere 5830 harvester with an on-board kernel processor. In the second experiment, Pioneer hybrids 3845 and Quanta were harvested at one-third milkline, two-thirds milkline, and blackline stages of maturity with and without mechanical processing. The theoretical length-of-cut was 12.7 mm. Total tract DM and OM digestibilities were lower for cows fed diets containing processed corn silage in experiment 1, and tended to be lower for cows fed diets containing unprocessed corn silage in experiment 2. Ruminal acetate concentrations were greater and ruminal propionate concentrations were lower 2 and 6 h after feeding for cows fed diets containing corn silage harvested at physiological maturity in experiment 2. This was due to decreased digestion of starch at advanced maturities in experiment 2. Ruminal pH tended to decline rapidly after feeding for cows fed hybrid Quanta (2 h) compared to hybrid 3845 (5 h) corn silage based diets. Ruminal acetate concentrations decreased and ruminal propionate concentrations increased 2 and 6 h after feeding for cows fed diets containing hybrid Quanta corn silage compared to hybrid 3845 corn silage. This was related to a greater starch concentration in the corn silage, greater starch intake, and increased rate of starch digestion for cows fed hybrid Quanta corn silage-based diets. Microbial nitrogen flow was lower and feed nitrogen flow was greater for cows fed diets containing hybrid Quanta corn silage. The lower microbial nitrogen flow was due to lower microbial nitrogen concentration and nonammonia nitrogen flow to the duodenum. Milk fat and protein concentrations had a strong quadratic relationship with forage NDF intake as a percentage of body weight. When forage NDF intake as a percentage of body weight dropped below 0.70%, there was a rapid decline in milk fat and protein concentrations.

Animals↗

Prolactin, interleukin-2 and FGF-2 stimulate expression, nuclear distribution and DNA-binding of rat homolog of pombe Cdc5 in Nb2 T lymphoma cells.

Pombe and human Cdc5 have been implicated in G2/M progression, but recently Cdc5 was identified as a component of a multiprotein complex essential for pre-mRNA splicing. We have previously isolated a prolactin (PRL)-inducible partial cDNA (1907 bp) encoding rat Cdc5. In the present study, the full length rCdc5 sequence (2847 bp) was obtained by 5'-RACE and cytokine regulation of Cdc5 expression was examined. PRL and interleukin-2 (IL2) act as mitogens in Nb2 T-lymphoma cells. Fibroblast growth factor (FGF-2) is not mitogenic in Nb2 cells but inhibits apoptosis of PRL-deprived cells. This study showed that PRL, IL-2 and FGF-2 rapidly increased Nb2 Cdc5 expression (3.4 kb mRNA) to reach 2-3-fold above controls at 4 h, and Cdc5 mRNA levels remained elevated at 24 h. There was a corresponding 2-3-fold increase in Cdc5 protein (105 kDa) levels at 24 h. Immunoblotting and fluorescent confocal microscopy showed predominant nuclear/perinuclear Cdc5 in quiescent Nb2 cells. PRL or FGF-2 treatment transiently increased nuclear Cdc5-specific immunofluorescence at 4 h but IL-2 gave maximal nuclear accumulation of Cdc5 at 24 h. The deduced rCdc5 protein has approximately 98% amino acid identity with human Cdc5. Like other Cdc5 family members, the N-terminus of rCdc5 contains two repeats of a DNA-binding domain found in a-, b- and c-Myb. Gel shift assays using (32)P-labeled Myb consensus oligonucleotides revealed two Myb-specific DNA-protein complexes in Nb2 nuclear extracts. Formation of both complexes was increased by PRL or FGF-2 at 1-5 and at 20 h and was partially inhibited by anti-Myb or anti-Cdc5 antibodies. In summary, rapid activation of Cdc5 in response to mitogenic and non-mitogenic stimuli suggests a complex role for Cdc5 in cellular regulation and this may not be restricted to mitotic entry or G2/M progression as previously supposed.

Animals↗

Consumption of watercress fails to alter coumarin metabolism in humans.

Watercress is an excellent source of phenethyl isothiocyanate (PEITC), an effective inhibitor of nitrosamine carcinogenesis in rodents. The mechanism of inhibition is believed to be due in part to inhibition of cytochrome P450 (P450) enzymes. P450 2A6 is a catalyst for the metabolic activation of several nitrosamines. In this study, we investigated the effect of watercress consumption on coumarin 7-hydroxylation, a P450 2A6-specific reaction, in a group of 15 nonsmoking, healthy volunteers. The urinary excretion of 7-hydroxycoumarin (7OHC) was determined. For 6 of the 15 subjects, watercress consumption decreased the amount of 7OHC excreted in the first 2 h following coumarin administration. However, the mean 0- to 2-h excretion of 7OHC for all 15 subjects was not significantly lowered by the consumption of watercress, 2.8 +/- 0.78 versus 3.1 +/- 0.53 mg (+/-S.D.). The mean 7OHC excreted from 2 to 4 h (1.1 +/- 0.50 mg) was significantly higher (P = 0.027) during watercress consumption than before (0.77 +/- 0.22 mg), suggesting a delay in coumarin metabolism. Total excretion of 7OHC was unaffected by watercress consumption. Therefore, under the conditions of our study, PEITC and other constituents of watercress had at most a marginal inhibitory effect on P450 2A6-catalyzed coumarin 7-hydroxylation.

Adult↗

Diamino benzo[b]thiophene derivatives as a novel class of active site directed thrombin inhibitors. 5. Potency, efficacy, and pharmacokinetic properties of modified C-3 side chain derivatives.

A systematic investigation of the structure-activity relationships of the C-3 side chain of the screening hit 1a led to the identification of the potent thrombin inhibitors 23c, 28c, and 31c. Their activities (1240, 903, and 1271 x 10(6) L/mol, respectively) represent 2200- and 2900-fold increases in potency over the starting lead 1a. This activity enhancement was accomplished with an increase of thrombin selectivity. The in vitro anticoagulant profiles of derivatives 28c and 31c were determined, and they compare favorably with the clinical agent H-R-1-[4aS, 8aS]perhydroisoquinolyl-prolyl-arginyl aldehyde (D-Piq-Pro-Arg-H; 32). The more potent members of this series have been studied in an arterial/venous shunt (AV shunt) model of thrombosis and were found to be efficacious in reducing clot formation. However, their efficacy is currently limited by their rapid and extensive distribution following administration.

Animals↗

Optimization of a fluoroscope to reduce radiation exposure in pediatric imaging.

BACKGROUND: A new children's hospital provided the impetus to investigate radiation dose and image quality in a fluoroscope that was specially engineered for pediatric fluoroscopy. Radiation protection management recommends radiation exposures that are as low as reasonably achievable, while still maintaining diagnostic image quality. OBJECTIVES: To obtain comparative phantom imaging data on radiation exposure and image quality from a newly installed fluoroscope before and after optimization for pediatric imaging. MATERIALS AND METHODS: Images were acquired from various thickness phantoms, simulating differing patient sizes. The images were evaluated for visualization of high- and low-contrast objects and for radiation exposure. Effects due to use of the image intensifier anti-scatter grid were also investigated. RESULTS: The optimization of the new fluoroscope for pediatric operation reduced radiation exposure by about 50% (compared to the originally installed fluoroscope), with very little loss of image quality. Pulsed fluoroscopy was able to lower radiation dose to less than 10% of continuous fluoroscopy, while still maintaining acceptable phantom image quality. CONCLUSION: Radiation exposure in pediatric fluoroscopy can be reduced to values well below the exposure settings that are typically found on unoptimized fluoroscopes. Pulsed fluoroscopy is considered a requisite for optimal pediatric fluoroscopy.

Adult↗

Development of a nursing management practice atlas. Part 2, Variation in use of nursing and financial resources.

Developing mechanisms for making benchmark comparisons among hospital organization is a challenge that has been embraced by nurse executives. A methodologic approach for ensuring data congruency when using available secondary data bases for making benchmark comparisons was detailed in part one (July/August) of this two-part series. This second article analyzes nursing management data using a set of nursing and financial resource variables identified by senior nurse executives of the hospital sites involved in this study.

Benchmarking↗

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

Data were collected from 10 experiments with duodenally cannulated Holstein dairy cows (271 combinations of cow and period) to evaluate the relationship between milk purine metabolites and microbial nitrogen flow. Experiments evaluated the effects of dietary factors on microbial N production and included: 1) supplemental sources of ruminally protected amino acids; 2) grass silage treated with fibrolytic enzymes; 3) bacterial inoculation of corn silage; 4) a comparison of corn and barley grain; 5) ruminal starch availability as affected by corn silages of varying maturity; and, 6) ruminal starch availability as affected by corn silages harvested at varying chop length and with or without mechanical processing. The coefficient of determination for individual experiments for the relationship between microbial nitrogen flow and allantoin or uric acid excretion in milk ranged from -0.01 to 0.77 and -0.06 to 0.22 respectively. Across all experiments, the coefficients of determination between microbial nitrogen flow and allantoin or uric acid excretion in milk were r2 = 0.09 and 0.01 respectively. Milk allantoin output was used to develop a prediction equation estimating microbial nitrogen flow to the duodenum. The greatest predictive value (r2 = 0.25) relationship was observed across non-bST experiments and included the factors of allantoin excretion, milk yield, dry matter intake, and days in milk. When milk yield was included in the model, the predictive value improved (r2 = 0.44). Based on these data, milk uric acid excretion alone cannot be used to predict microbial N production accurately [corrected].

Allantoin↗

Novobiocin-induced VP-16 accumulation and MRP expression in human leukemia and ovarian carcinoma cells.

We have previously reported that novobiocin potentiates the cytotoxic activity of etoposide (VP-16) and teniposide (VM-26) in a number of experimental tumor cell lines by inhibition of the efflux of the epipodophyllotoxins by an ATP-requiring transporter. In leukemia cells from 12/19 patients and in ovarian carcinoma cells from 2/4 patients, novobiocin, in a concentration range of 150-1000 microM, increased the intracellular accumulation of VP-16 by 30-250% by inhibiting its efflux. Novobiocin did not significantly increase the intracellular concentration of VP-16 in human mononuclear bone marrow cells from two individuals with normal bone marrow, suggesting that it might be possible to selectively modulate the intracellular accumulation of the epipodophyllotoxin in tumor cells relative to normal hematopoietic tissue. Previous findings from our laboratory have provided evidence that the membrane transporter for VP-16 which is inhibited by novobiocin is distinct from the P-glycoprotein. The expression of MRP, measured by immunoblotting, was variable in novobiocin-responsive and non-responsive leukemia cells, indicating that no direct relationship existed between the modulatory activity of novobiocin on the transport of VP-16 and the expression of the MRP gene. The findings indicate that the novobiocin-sensitive VP-16 transporter is (i) present in high frequency in leukemia and ovarian carcinoma cells, and (ii) probably not the P-glycoprotein or MRP.

ATP Binding Cassette Transporter, Subfamily B, Mem↗