Shawn S. Donkin

Gluconeogenesis in dairy cows: the secret of making sweet milk from sour dough.

Gluconeogenesis is a crucial process to support glucose homeostasis when nutritional supply with glucose is insufficient. Because ingested carbohydrates are efficiently fermented to short‐chain fatty acids in the rumen, ruminants are required to meet the largest part of their glucose demand by de novo genesis after weaning. The qualitative difference to nonruminant species is that propionate originating from ruminal metabolism is the major substrate for gluconeogenesis. Disposal of propionate into gluconeogenesis via propionyl‐CoA carboxylase, methylmalonyl‐CoA mutase, and the cytosolic form of phosphoenolpyruvate carboxykinase (PEPCK) has a high metabolic priority and continues even if glucose is exogenously supplied. Gluconeogenesis is regulated at the transcriptional and several posttranscriptional levels and is under hormonal control (primarily insulin, glucagon, and growth hormone). Transcriptional regulation is relevant for regulating precursor entry into gluconeogenesis (propionate, alanine and other amino acids, lactate, and glycerol). Promoters of the bovine pyruvate carboxylase (PC) and PEPCK genes are directly controlled by metabolic products. The final steps decisive for glucose release (fructose 1,6‐bisphosphatase and glucose 6‐phosphatase) appear to be highly dependent on posttranscriptional regulation according to actual glucose status. Glucogenic precursor entry, together with hepatic glycogen dynamics, is mostly sufficient to meet the needs for hepatic glucose output except in high‐producing dairy cows during the transition from the dry period to peak lactation. Lactating cows adapt to the increased glucose requirement for lactose production by mobilization of endogenous glucogenic substrates and increased hepatic PC expression. If these adaptations fail, lipid metabolism may be altered leading to fatty liver and ketosis. Increasing feed intake and provision of glucogenic precursors from the diet are important to ameliorate these disturbances. An improved understanding of the complex mechanisms underlying gluconeogenesis may further improve our options to enhance the postpartum health status of dairy cows.

Vitamin D: emerging new roles in insulin sensitivity.

The growing incidence of prediabetes and clinical type 2 diabetes, in part characterised by insulin resistance, is a critical health problem with consequent devastating personal and health-care costs. Vitamin D status, assessed by serum 25-hydroxyvitamin D levels, is inversely associated with diabetes in epidemiological studies. Several clinical intervention studies also support that vitamin D, or its active metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D), improves insulin sensitivity, even in subjects with glucose metabolism parameters classified within normal ranges. The mechanisms proposed which may underlie this effect include potential relationships with improvements in lean mass, regulation of insulin release, altered insulin receptor expression and specific effects on insulin action. These actions may be mediated by systemic or local production of 1,25(OH)2D or by suppression of parathyroid hormone, which may function to negatively affect insulin sensitivity. Thus, substantial evidence supports a relationship between vitamin D status and insulin sensitivity; however, the underlying mechanisms require further exploration.

1α,25-Dihydroxyvitamin D hydroxylase in adipocytes ☆

High vitamin D intake is associated with reduced insulin resistance. Expression of extra-renal 1alpha,25-dihydroxyvitamin D hydroxylase (1alpha-hydroxylase) has been reported in several tissues and contributes to local synthesis of 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D) from the substrate 25-hydroxyvitamin D (25OHD). Expression and dietary regulation of 1alpha-hydroxylase in tissues associated with energy metabolism, including adipose tissue, has not been assessed. Male Wistar rats were fed a high calcium (1.5%) and high vitamin D (10,000IU/kg) or a low calcium (0.25%), low vitamin D (400IU/kg) with either a high fat (40% energy) or high sucrose (66% energy) dietary background for 14 weeks. Expression of 1alpha-hydroxylase, assessed by real time PCR, was detected in adipose tissue and did not differ with dietary level of calcium and vitamin D. 1alpha-Hydroxylase mRNA was also detected in 3T3-L1 preadipocytes and 25OHD treatment at 10nM levels induced 1,25(OH)(2)D responsive gene, CYP24, and this response was reduced in the presence of the p450 inhibitor, ketoconazole. In addition, (3)H 25OHD was converted to (3)H 1,25(OH)(2)D in intact 3T3-L1 preadipocytes. Cumulatively, these results demonstrate that 1alpha-hydroxylase is expressed in adipose tissue and is functional in cultured adipocytes. Thus, the capacity for local production may play a role in regulating adipocyte growth and metabolism.

Feeding value of glycerol as a replacement for corn grain in rations fed to lactating dairy cows.

Growth of the corn ethanol industry has created a need for alternatives to corn for lactating dairy cows. Concurrent expansion in soydiesel production is expected to increase availability and promote favorable pricing for glycerol, a primary co-product material. The objective of this study was to determine the feeding value of glycerol as a replacement for corn in diets fed to lactating dairy cattle. Sixty lactating Holstein cows housed in individual tie stalls were fed a base diet consisting of corn silage, legume forages, corn grain, soyhulls, roasted soybeans, and protein supplements. After a 2-wk acclimation period, cows were fed diets containing 0, 5, 10, or 15% refined glycerol for 56 d. Cows were milked twice daily and weekly milk samples were collected. Milk production was 36.3, 37.2, 37.9, and 36.2 +/- 1.6 kg/d and feed intake was 23.8, 24.6, 24.8, and 24.0 +/- 0.7 kg/d for 0, 5, 10, and 15% glycerol treatments, respectively, and did not differ except for a modest reduction in feed intake during the first 7 d of the trial for 15% glycerol (treatment x time effect). Milk composition was not altered by glycerol feeding except that milk urea nitrogen was decreased from 12.5 +/- 0.4 to 10.2 +/- 0.4 mg/dL with glycerol addition. Cows fed diets containing 10 and 15% glycerol gained more weight than those fed rations containing 0 or 5% glycerol but body condition scores did not differ with glycerol feeding. The data indicate that glycerol is a suitable replacement for corn grain in diets for lactating dairy cattle and that it may be included in rations to a level of at least 15% of dry matter without adverse effects on milk production or milk composition.

A self referencing platinum nanoparticle decorated enzyme-based microbiosensor for real time measurement of physiological glucose transport

Glucose is the central molecule in many biochemical pathways, and numerous approaches have been developed for fabricating micro biosensors designed to measure glucose concentration in/near cells and/or tissues. An inherent problem for microsensors used in physiological studies is a low signal-to-noise ratio, which is further complicated by concentration drift due to the metabolic activity of cells. A microsensor technique designed to filter extraneous electrical noise and provide direct quantification of active membrane transport is known as self-referencing. Self-referencing involves oscillation of a single microsensor via computer-controlled stepper motors within a stable gradient formed near cells/tissues (i.e., within the concentration boundary layer). The non-invasive technique provides direct measurement of trans-membrane (or trans-tissue) analyte flux. A glucose micro biosensor was fabricated using deposition of nanomaterials (platinum black, multiwalled carbon nanotubes, Nafion) and glucose oxidase on a platinum/iridium microelectrode. The highly sensitive/selective biosensor was used in the self-referencing modality for cell/tissue physiological transport studies. Detailed analysis of signal drift/noise filtering via phase sensitive detection (including a post-measurement analytical technique) are provided. Using this highly sensitive technique, physiological glucose uptake is demonstrated in a wide range of metabolic and pharmacological studies. Use of this technique is demonstrated for cancer cell physiology, bioenergetics, diabetes, and microbial biofilm physiology. This robust and versatile biosensor technique will provide much insight into biological transport in biomedical, environmental, and agricultural research applications.

Glycerol from biodiesel production: the new corn for dairy cattle

Glycerol, also known as glycerin, is a colorless, odorless, hygroscopic, and sweet-tasting viscous liquid. It is a sugar alcohol with high solubility index in water and has a wide range of applications in the food, pharmaceutical, and cosmetic industries. The use of glycerol in diets for dairy cattle is not novel; however, this interest has been renewed due to the increased availability and favorable pricing of glycerol as a consequence of recent growth in the biofuels industry. Experimental evidence supports the use of glycerol as a transition cow therapy but feeding rates are low, ranging from 5 to 8 % of the diet DM. There is a paucity of research that examines the use of glycerol as a macro-ingredient in rations for lactating dairy cows. Most reports indicate a lack of effect of addition of glycerol to the diet when it replaces corn or corn starch. Recent feeding experiments with lactating dairy cows indicate replacing corn with glycerol to a level of 15% of the ration DM does not adversely effect milk production or composition. Milk production was 37.0, 36.9, 37.3, 36.4 ± 0.6 kg/d and feed intake was 24.0, 24.5, 24.6, 24.1 ± 0.5 kg/d for 0, 5, 10 and 15% glycerol treatments respectively and did not differ (P > 0.05) except for a modest reduction in feed intake during the first 7 days for the 15% glycerol treatment. Glycerol fed to dairy cattle is fermented to volatile fatty acids in the rumen and early reports indicated that glycerol is almost entirely fermented to propionate. In vitro data indicates glycerol fermentation increases the production of propionate and butyrate at the expense of acetate. Rumen microbes appear to adapt to glycerol feeding and consequently, cows fed glycerol also require an adaptation period to glycerol inclusion. Debate exists regarding the fate of glycerol in the rumen and although most reports suggest that glycerol is largely fermented in the rumen, the extent of rumen digestion may depend on level of inclusion in the diet. Data are lacking regarding the rates of rumen fermentation of glycerol at intake levels for high producing dairy cattle. Current data indicates that glycerol can be included in diets fed to dairy cattle at macro ingredient levels; however, additional information is needed to permit a full appreciation of the feeding value of glycerol and the resulting impact on cow health and productivity.

Parathyroid Hormone Suppresses Insulin Signaling in Adipocytes

Previous reports suggest that parathyroid hormone (PTH) is associated with insulin resistance. This research investigated the effects of PTH on insulin signaling in differentiated 3T3-L1 adipocytes. PTH (10 nM, 24 h) treatment induced a reduction in insulin-stimulated glucose uptake, AKT activity (phosphorylated AKT/total AKT protein expression) and a decrease in GLUT4 and IRS-1 protein expression compared to vehicle treated controls in differentiated adipocytes. PTH treatment also induced increased phosphorylation of IRS-1 on serine 307, which suppresses insulin signaling. In addition, treatment of cells with adenyl cyclase inhibitor SQ52236 ameliorated the effects of PTH on insulin-stimulated glucose uptake, whereas inhibition of phospholipase C alpha (U73122) did not significantly alter the effects of PTH. Thus, PTH treatment of differentiated 3T3-L1 adipocytes suppresses insulin-stimulated glucose uptake and insulin signaling via cAMP pathway, potentially through the phosphorylation of IRS-1 at serine 307.

Dietary Intervention with Vitamin D, Calcium and Whey Protein Reduced Fat Mass and Increased Lean Mass in Rats

The aim of the current study was to determine the effects and the mechanisms of inclusion of dietary whey protein, high calcium, and high vitamin D intake with either a high-sucrose or high-fat base diets on body composition of rodents. Male Wistar rats were assigned to either no whey protein, suboptimal calcium (0.25%), and vitamin D (400 IU/kg) diet (LD), or a diet containing whey protein, high calcium (1.5%), and vitamin D (10 000 IU/kg) diet (HD), and either high-fat (40% of energy) or high-sucrose (60%) base diets for 13 weeks. Liver tissue homogenates were used to determine [(14)C]glucose and [(14)C]palmitate oxidation. mRNA expression of enzymes related to energy metabolism in liver, adipose, and muscle, as well as regulators of muscle mass and insulin receptor was assessed. The results demonstrated that there was reduced accumulation of body fat mass (P = .01) and greater lean mass (P = .03) for the HD- compared to LD-fed group regardless of the background diet. There were no consistent differences between the LD and HD groups across background diets in substrate oxidation and mRNA expression for enzymes measured that regulate energy metabolism, myostatin, or muscle vascular endothelial growth factor. However, there was an increase in insulin receptor mRNA expression in muscle in the HD compared to the LD groups. In conclusion, elevated whey protein, calcium, and vitamin D intake resulted in reduced accumulation of body fat mass and increased lean mass, with a commensurate increase in insulin receptor expression, regardless of the level of calories from fat or sucrose.

Feeding conjugated linoleic acid partially recovers carcass quality in pigs fed dried corn distillers grains with solubles

Dried corn distillers grains with solubles (DDGS) fed to swine may adversely affect carcass quality due to the high concentration of unsaturated fat. Feeding CLA enhances pork quality when unsaturated fat is contained in the diet. The effects of CLA on growth and pork quality were evaluated in pigs fed DDGS. Diets containing 0, 20, or 40% DDGS were fed to pigs beginning 30 d before slaughter. At 10 d before slaughter, one-half of each DDGS treatment group was fed 0.6% CLA or 1% choice white grease. Carcass data, liver- and backfat-samples were collected at slaughter. Longissimus muscle area, 10th-rib back-fat depth, last rib midline backfat depth, LM color, marbling, firmness and drip loss, and bacon collagen content were not altered by DDGS or CLA. Outer layer backfat iodine values were increased (P <or= 0.05) with DDGS feeding and were 65.07, 69.75, and 74.25 for 0, 20, and 40% DDGS, respectively. Addition of CLA decreased (P <or= 0.05) outer layer backfat iodine values from 71.11 to 68.31. Diets containing DDGS decreased (P <or= 0.05) percent lean tissue contained in bacon from 48% for controls to 38% for pigs fed 40%. Abundance of fatty acid synthase, carnitine palmitoyl transferase Ia, acetyl-CoA-carboxylase, stearoyl-CoA desaturase, and glycerol-3-phosphate dehydrogenase mRNA in adipose or liver were not different (P > 0.05) for pigs fed DDGS. Feeding CLA decreased (P <or= 0.05) the Delta(9) de-saturase index in adipose tissue. The data indicate that decreased carcass firmness with DDGS feeding is not reflected by changes in lipogenic gene expression. Feeding 20% or more DDGS to finishing swine decreases bacon leanness, but inclusion of 0.6% CLA in the finishing diet can partially reverse these effects.

Effects of temperature stress on growth performance and bacon quality in grow-finish pigs housed at two densities

Managing stressors is essential for optimizing pig growth performance. To determine the effects of temperature and space allocation on growth performance and carcass characteristics, pigs were housed within their thermoneutral zone, at 23.9 degrees C, or above their thermoneutral zone, at 32.2 degrees C, and were provided either 0.66 or 0.93 m(2)/pig for the final 35 d of the grow-finish period. Individual BW were recorded on d 1, 10, 20, and 30. At slaughter, carcass measurements and samples of backfat and belly fat were collected. Final BW was decreased (P < or = 0.05) from 113 to 103 kg for pigs housed at 32.2 degrees C. The ADG was reduced (P < 0.05) for pigs housed at 32.2 degrees C (0.89 vs. 0.54 kg/d), as was G:F (0.28 vs. 0.24). Housing at 0.66 m(2)/pig resulted in pigs that were lighter (P < or = 0.05), at 106 compared with 110 kg, as a result of decreased (P < or = 0.05) ADG (0.78 to 0.65 kg/d) and decreased (P < or = 0.05) G:F (0.275 to 0.255) compared with pigs housed at 0.93 m(2)/pig. Pigs housed at a greater spatial allocation had elevated (P < or = 0.05) ADFI. The interaction of housing at 32.2 degrees C and decreasing spatial allocation increased (P < or = 0.05) the adipose iodine value from 66.8 to 70.4, decreased (P < or = 0.05) the saturated:unsaturated fatty acids ratio from 0.59 to 0.56, and increased (P < or = 0.05) the n-6:n-3 from 23.56 to 25.27. Decreased spatial allocation resulted in decreased (P < or = 0.05) belly weights. Although increased temperature did not affect belly weight, the 32.2 degrees C pigs had decreased (P < or = 0.05) raw and cooked slice weights, increased (P < or = 0.05) percentage lean of bacon, increased (P < or = 0.05) lean:fat ratio of bacon slices, increased (P < or = 0.05) raw slice scores, and increased (P < or = 0.05) quantity of collagen in belly fat. Some of these changes may have resulted from changes in lipid metabolism. Increasing spatial allocation in the 32.2 degrees C pigs decreased fatty acid synthase (P = 0.03) and stearoyl-CoA desaturase- 1 (P = 0.08) mRNA expression in adipose tissue. The results from this study demonstrated decreased growth, carcass lipid quality, and bacon quality in pigs housed at temperatures above the thermoneutral zone; however, increasing the spatial allocation for housing may be a means to ameliorate the negative effects of temperature stress.