Donald C. Beitz

Strong relationships between mediators of the acute phase response and fatty liver in dairy cows

The objective of this study was to investigate the relationship between activation of acute phase response and fatty liver in transition dairy cows. Fatty liver was induced in dairy cows by feeding 8 kg of cracked corn 1 mo before the expected day of parturition. Liver and blood samples were obtained at days -4, 3, 8, 12, 14, 22, 27, and 36 postpartum. Cows that developed fatty liver (n = 4) reached peak total lipids in the liver at day 12 postpartum with 11.4% (wet wt.) compared with 6.6% in control cows (n = 4). Cows with fatty liver had greater plasma tumor necrosis factor-alpha (TNF-α), nonesterified fatty acids (NEFA), and lower lactate concentrations than did control cows at day -4. During highest concentrations of total lipids in the liver, for at least one time-point, fatty-liver cows had greater concentrations of plasma serum amyloid A (SAA), haptoglobin, and NEFA and lower concentrations of calcitonin gene-related peptide (CGRP), prostaglandin E2 (PGE2), cortisol, and TNF-α than did control cows...

Resveratrol promotes atherosclerosis in hypercholesterolemic rabbits

The hypothesis was tested that resveratrol, a compound in red wine, would inhibit atherosclerotic development in rabbits fed 0.5% cholesterol for 60 days. Rabbits were supplemented with or without oral resveratrol. During the study, body weights and food consumption were similar for the two groups. The lack of differences between liver weights and a series of serum parameters indicative of liver disease suggest that liver function was similar in the two groups. The diet produced hypercholesterolemia in both groups, but no differences in lipoprotein-cholesterol concentrations. The electrophoretic mobility of plasma low-density lipoprotein (LDL) and plasma LDL after induced oxidation also was not different between the groups. Staining of atherosclerotic lesions in the control and resveratrol-treated groups revealed that the resveratrol-treated rabbits had significantly more aortic surface area covered by atherosclerotic lesions (P < 0.02). Therefore, resveratrol promoted atherosclerotic development, rather than protect against it, by a mechanism that is independent of observed differences in gross animal health, liver function, plasma cholesterol concentrations, or LDL oxidative status.

The inhibitory effects of β-amyloid on glutamate and glucose uptakes by cultured astrocytes

beta-Amyloid is the primary protein component of neuritic plaques, which are degenerative foci in brains of patients with Alzheimers disease (AD). The effects of this naturally occurring beta-amyloid on the cells of the central nervous system have not been completely understood. beta-Amyloid increases the vulnerability of cultured neurons to glutamate-induced excitotoxic damage. Because astrocytes play a key role in uptake of extracellular glutamate and glutamate uptake is ATP-dependent, we studied the effect of beta25-35 on glutamate and glucose uptake in cultured hippocampal astrocytes following 7 days of exposure to beta25-35. Astrocytic glutamate uptake was studied at 1, 5, 10, 15, 20, and 60 min following the addition of [3H]glutamate (5 nM) to the culture media, and astrocytic glucose uptake was assessed at 60 min after the addition of [14C]glucose (600 and 640 nM) to the media. Glutamate uptake by control astrocytes was time-dependent. Astrocytes exposed to beta25-35, however, showed significantly lower glutamate uptake at all sampling times. Similarly, [14C]glucose uptake by astrocytes was inhibited by beta25-35. When glucose uptake was blocked by phloretin (10 mM), astrocytic [3H]glutamate uptake was also blocked, suggesting that the inhibitory effect of beta-amyloid on glutamate uptake is caused by diminished glucose uptake. Thus, our present study suggests a possible link between two proposed mechanisms of pathogenesis of the Alzheimers disease: glutamate neurotoxicity and global defect in cerebral energy metabolism.

1,25-Dihydroxyvitamin D3 receptor in bovine thymus gland

Abstract 1,25-Dihydroxyvitamin D 3 [1,25-(OH) 2 D 3 ] receptor was characterized after partial purification of thymus cytosol by ammonium sulfate fractionation. The 1,25-(OH) 2 D 3 receptor sediments at 3.7S in 5–20% sucrose gradients. The binding of 1,25-(OH) 2 D 3 in thymic cytosol was a saturable process with high affinity (Kd = 0.12−0.48 nM) at 4°C. Competition for 1,25-(OH) 2 [ 3 H]D 3 receptor by nonradioactive analogs demonstrated the affinities of these analogs to be in order; 1,25-(OH) 2 D 3 = 1,24R,25-(OH) 3 D 3 = 1,25S,26-(OH) 3 D 3 = 1,25R,26-(OH) 3 D 3 > 1,25-(OH) 2 D 3 -26,23 lactone > 25-OHD 3 > 23R,25-(OH) 2 D 3 > 24R,25-(OH) 2 D 3 > 23S,25-(OH) 2 D 3 ⋙ 25-OHD 3 -26,23 lactone. The receptor bound to DNA cellulose columns in low salt buffer and eluted as a single peak at 0.21 M KCl. These findings provide evidence that the thymus possesses a 1,25-(OH) 2 D 3 receptor with properties indistinguishable from 1,25-(OH) 2 D 3 receptors in other tissues.

The effect of mitochondrial DNA on milk production and health of dairy cattle

Maternal lineage effects, probably indicative of mitochondrial DNA (mtDNA) differences, may be important for milk production and reproductive success in dairy cattle (Bos taurus). Sequence variation of mtDNA in 36 maternal lineages of dairy cattle was studied with animal models to assess effects on milk production and reproductive traits. Cattle within maternal lineages defined by registered pedigrees were assumed to be uniform for the nucleotide sequences examined. Sequence polymorphisms of bovine mtDNA were shown to be associated with milk production, reproduction, and health costs incurred. One particular base-pair substitution was associated with additional production of 842 kg milk and 37 kg milk fat per cow per lactation. Another single base-pair substitution was associated with a decrease of 36 days and one unsuccessful breeding between successive calvings. Effects of this size are economically important and have broad implications in genetic selection of dairy cattle.

Estimation of relationships between mineral concentration and fatty acid composition of longissimus muscle and beef palatability traits

The objective of this study was to determine the influence of beef LM nutrient components on beef palatability traits and evaluate the impact of USDA quality grade on beef palatability. Longissimus muscle samples from related Angus cattle (n = 1,737) were obtained and fabricated into steaks for trained sensory panel, Warner-Bratzler shear force (WBSF), lipid oxidation measured by thiobarbituric acid reactive substances (TBARS), fatty acid, and mineral composition analysis. Pearson phenotypic correlations were obtained by the correlation procedure of SAS. Beef palatability data were analyzed by the GLM procedure of SAS with USDA quality grade as the main effect. Specific mineral concentrations did not demonstrate strong correlations with WBSF or sensory traits (r = -0.14 to 0.16). However, minerals appeared to have a stronger relationship with flavor; all minerals evaluated except Ca and Mn were positively correlated (P < 0.05) with beef flavor. Stearic acid (C18:0), C18:2, C20:4, and PUFA were negatively correlated (P < 0.05) with all 3 panelist tenderness traits (r = -0.09 to -0.22) and were positively correlated (P < 0.05) with WBSF (r = 0.09 to 0.15). The MUFA were positively correlated (P < 0.05) with panelist tenderness ratings (r = 0.07 to 0.10) and negatively associated (P < 0.05) with WBSF (r = -0.11). The strongest correlations with juiciness were negative relationships (P < 0.05) with C18:2, C18:3, C20:4, and PUFA (r = -0.08 to -0.20). Correlations with beef flavor were weak, but the strongest was a positive relationship with MUFA (r = 0.13). Quality grade affected (P < 0.05) WBSF, TBARS, and all trained sensory panel traits, except livery/metallic flavor. As quality grade increased, steaks were more tender (P < 0.05), as evidenced by both WBSF and sensory panel tenderness ratings. Prime steaks were rated juiciest (P < 0.05) by panelists, whereas Select and Low Choice were similarly rated below Top Choice for sustained juiciness. Quality grade influenced (P < 0.05) beef flavor, but not in a linear fashion. Although there were significant correlations, these results indicate tenderness, juiciness, and flavor are not strongly influenced by individual nutrient components in beef LM. Furthermore, the positive linear relationships between USDA quality grade and beef palatability traits suggest quality grade is still one of the most valuable tools available to predict beef tenderness.

Mechanism of cholesterol reduction to coprostanol by Eubacterium coprostanoligenes ATCC 51222

The mechanism of reduction of cholesterol to coprostanol by Eubacterium coprostanoligenes ATCC 51222 was studied by incubating the bacterium with a mixture of alpha- and beta-isomers of [4-3H,4-14C]cholesterol. Coprostanol, isolated after incubation of [4-3H,4-14C]cholesterol in a growth medium under anaerobic conditions, retained 97% of the tritium originally present in cholesterol. The majority of this tritium (64%), however, was in the C-6 position in coprostanol, which showed that the conversion of cholesterol into coprostanol by E. coprostanoligenes involved the intermediate formation of 4-cholesten-3-one followed by the reduction of the latter to coprostanol. In resting cell assays in which washed bacterial cells were incubated with micellar cholesterol in phosphate buffer at 37 degrees C, both 4-cholesten-3-one and coprostanone were produced in addition to coprostanol. Furthermore, 5-cholesten-3-one, 4-cholesten-3-one, and coprostanone were converted efficiently to coprostanol by E. coprostanoligenes. These results support the hypothesis that the major pathway for reduction of cholesterol by E. coprostanoligenes involves the intermediate formation of 4-cholesten-3-one followed by reduction of the latter to coprostanol through coprostanone as an intermediate.

1,25-Dihydroxyvitamin D3 inhibits secretion of interferon-γ by mitogen- and antigen-stimulated bovine mononuclear leukocytes☆

1,25-Dihydroxyvitamin D3 (1,25(OH)2D3), the biologically active metabolite of vitamin D, and delta 22-26-F3-1,25-dihydroxyvitamin D3 (delta 22-26-F3-1,25(OH)2D3), a synthetic analog with a high affinity for the vitamin D receptor, significantly inhibited interferon-gamma (IFN-gamma) secretion in 24- and 48-h cultures of pokeweed mitogen (PWM) and ovalbumin (OVA) stimulated peripheral blood mononuclear leukocyte (MNL) from adult, OVA-sensitized dairy cattle. Vitamin D-induced inhibition of IFN-gamma production was most pronounced in MNL cultures supplemented with 1,25(OH)2D3 at 1.0 nM or more, a concentration equal to or exceeding that in plasma of cows with clinical hypocalcemia. Secreted IFN-gamma was undetectable in all resting MNL cultures. Ultra-low concentrations (0.0001, 0.001, and 0.01 nM) of 1,25(OH)2D3 had no effect on IFN-gamma secretion by PWM-stimulated bovine MNL, unlike a previous study in other species demonstrating enhancement of IFN-gamma secretion at these concentrations. Preincubation of MNL with.

Short-term consumption of sucralose, a nonnutritive sweetener, is similar to water with regard to select markers of hunger signaling and short-term glucose homeostasis in women

Nonnutritive sweeteners have been used to lower the energy density of foods with the intention of affecting weight loss or weight maintenance. However, some epidemiological and animal evidence indicates an association between weight gain or insulin resistance and artificial sweetener consumption. In the present study, we hypothesized that the nonnutritive sweetener sucralose, a trichlorinated sucrose molecule, would elicit responses similar to water but different from sucrose and sucrose combined with sucralose on subjective and hormonal indications of hunger and short-term glucose homeostasis. Eight female volunteers (body mass index, 22.16 ± 1.71 kg/m(2); age, 21.75 ± 2.25 years) consumed sucrose and/or sucralose in water in a factorial design. Blood samples were taken at fasting and 30 and 60 minutes after treatment followed by a standardized breakfast across treatments, and blood samples were taken 30, 60, 90, and 120 minutes after breakfast. Plasma was analyzed for glucose, insulin, glucagon, triacylglycerols (TAG), and acylated ghrelin. Perceptions of hunger and other subjective measurements were assessed before each blood sample. No differences were detected in subjective responses, circulating triacylglycerol, or glucagon concentrations among treatments over time. Significant differences were observed in insulin, glucose, and acylated ghrelin concentrations over time only between sucrose-containing treatments and non-sucrose-containing treatments regardless of sucralose consumption. Therefore, sucralose may be a relatively inert nonnutritive sweetener with regard to hunger signaling and short-term glucose homeostasis.

A sensitive competitive protein binding assay for vitamin D in plasma.

A sensitive protein binding assay for vitamin D is described. The vitamin D3 was extracted from plasma with diethyl ether and methylene chloride. The lipid extract was purified in Sephadex LH-20 followed by Lipidex 5000 and finally by high pressure liquid chromatography on a Zorbax Sil column (0.79 x 25 cm) developed in 0.25:99.75 isopropanol: methylene chloride. The vitamin D fraction was collected and quantitated by competitive protein binding assay with a 1/50,000 dilution of sheep plasma in 0.05 M potassium phosphate buffer (pH 7.5) containing 0.01% gelatin. [H3]-25-Hydroxyvitamin D3 was used as a radioactive tracer in the assay. We found that under these conditions, sheep plasma had equal affinity for vitamin D2 and vitamin D3 and could detect as little as 0.1 ng of vitamin D. When rat, cow, or human plasma was substituted for the sheep plasma, the decline in sensitivity to vitamin D2 was fivefold to tenfold. With this assay, we found excellent agreement (r = 0.98) between the results obtained by competitive protein binding analysis and direct U.V. absorbance analysis by high pressure liquid chromatography.