Effect of algae or fish oil supplementation and porcine maternal stress on the adrenal transcriptome of male offspring fed a low-quality protein diet.

Offspring adrenal function may be negatively affected in utero by maternal stressors such as microbial infection. Maternal supplementation with immunomodulatory compounds such as omega-3 polyunsaturated fatty acids (n-3 PUFA) may help minimize the adverse effects of maternal stress on fetal hypothalamic-pituitary-adrenal development and improve offspring health. Presently, n-3 PUFA sources are primarily fish-based, but n-3 PUFA microalgae (AL) may be an alternative. Previously, it was determined that maternal AL or fish oil (FO) supplementation to sows, in addition to maternal stress induced by Escherichia coli lipopolysaccharide (LPS) challenge appeared to have a greater influence on the stress response of male offspring compared to females. To further elaborate on these findings, this study assessed the effects of maternal AL or FO supplementation combined with a maternal LPS challenge on adrenal gene expression in male offspring fed a nursery diet containing low-quality protein sources. Forty-eight sows were fed gestation diets starting on gestation day (gd) 75 containing either 3.12% AL, 3.1% FO, or a control diet containing 1.89% corn oil. On gd 112, half the sows in each treatment were administered 10 â€‹µg/kg LPS i.m. Piglets were weaned at 21 days of age onto a common low-quality plant-based protein diet, and one week after weaning, four piglets per sow were administered 40 â€‹µg/kg LPS i.m. Two hours later, the piglets were euthanized to obtain adrenal tissue, and total RNA was extracted to carry out transcriptome analysis using the Affymetrix GeneChip WT Plus assay and subsequent validation by real-time PCR. Analysis revealed that adrenal steroidogenesis, fatty acid metabolism and immune function were significantly influenced by maternal diet and stress. Increased expression of immune-related genes including lymphocyte antigen 96, TLR-2 and NF-κB suggests that maternal AL supplementation may increase offspring sensitivity to inflammation after weaning. Decreased expression of lymphocyte antigen 96 in male offspring from sows receiving maternal LPS challenge also suggests a possible role of maternal stress in diminishing the offspring immune response to immune stress challenge. Increased expression of the genes encoding the 11BHSD2 enzyme in offspring from sows fed FO may also reduce the magnitude of the stress response. These data provide insight to the immune and metabolic mechanisms that may be influenced by maternal diet and stress.

Supplementation of organic and inorganic selenium to late gestation and early lactation beef cows effect on progeny feedlot performance and carcass characteristics.

Angus × Simmental cows ( = 48, BW = 594 kg, BCS = 5.26, age = 2.7) pregnant with male fetuses were used to determine the effect of selenium source during the last 80 d of gestation and first 108 d of lactation on progeny feedlot performance. At 203 d of gestation, cows were blocked by BW, breed composition, and calf sire and randomly allotted to 1 of 3 treatments: no supplemental Se, 3 mg/d inorganic Se (sodium selenite), and 3 mg/d organic Se (Sel-plex). Maternal diets were formulated to contain 10.4% CP and 0.90 Mcal/kg NE during gestation and 12.1% CP and 1.01 Mcal/kg NE during lactation. Basal diets contained 0.07 and 0.11 mg/kg Se for gestation and lactation diets, respectively. Diets were fed daily as a total mixed ration, and no additional Se, 3 mg/d Se as sodium selenite, or 3 mg/d Se as Sel-Plex were top-dressed daily. Treatment diets were fed through 108 d postpartum (DPP). At 108 DPP cow-calf pairs were commingled until weaning at 210 DPP. At 28 d postweaning, steers ( = 47, BW = 301 kg) were placed in individual pens and fed a diet formulated to provide 13.9% CP and 1.24 Mcal/kg NE. No supplemental Se was fed; however, basal Se concentration was 0.10 mg/kg. The diet was delivered as a total mixed ration once daily. Steers were slaughtered at a target BW of 625 kg. Steers from cows supplemented with organic Se tended to enter the feedlot heavier ( = 0.06) than steers from cows supplemented with inorganic Se. There was no difference in ADG among treatments ( ≥ 0.73), but steers from organic Se cows tended to spend fewer days on feed compared to steers from inorganic Se cows ( = 0.09). Steers from organic Se cows had a greater overall DMI compared to steers from inorganic Se cows ( = 0.04), but there was no difference in overall G:F ( = 0.49). Dressing percentage was greater for steers from cows fed no Se compared with steers from cows fed either inorganic or organic Se ( = 0.03). Maternal Se source had no effect on HCW, back fat, percentage KPH, LM area, yield grade, marbling score, or quality grade distribution ( ≥ 0.17) of progeny. In conclusion, maternal supplementation with organic Se appears to have a long-term benefit on intake of steer progeny and may result in improvements in growth that could decrease days on feed.

Supplementation of organic and inorganic selenium to late gestation and early lactation beef cows effect on cow and preweaning calf performance.

Angus × Simmental cows ( = 48; BW = 595 ± 17.4 kg, BCS = 5.26 ± 0.05, and age = 2.3 ± 0.07 yr), pregnant with male fetuses, were used to determine the effect of Se source during the last 80 d of gestation and first 108 d of lactation on cow and calf performance. At 203 d in gestation, cows were blocked by BW, breed composition, age, and calf sire and randomly allotted to organic Se, inorganic Se, or no Se treatments. Diets contained corn silage, corn stover, haylage, dried distillers' grains with solubles, and minerals and were formulated to contain 10.4% CP and 0.90 Mcal/kg NEg during gestation and 12.1% CP and 1.01 Mcal/kg NEg during lactation. Diets were fed daily as a total mixed ration and none, 3 mg/d Se as sodium selenite, or 3 mg/d Se as Sel-Plex were top-dressed daily. At 68 d postpartum (DPP), milk production was calculated using the weigh-suckle-weigh procedure and a milk sample was collected to determine composition. At 108 DPP, cow-calf pairs were commingled until weaning at 210 DPP. Cow BW and BCS ( ≥ 0.56) did not differ between treatments at any time point during the study. Milk production, milk fat, and total solids ( ≥ 0.38) did not differ among treatments. Milk protein tended to increase in cows fed inorganic Se compared with cows fed organic Se ( = 0.07) and milk lactose tended to be greatest in cows fed organic Se ( = 0.10). Conception to AI and overall pregnancy rates did not differ between treatments ( ≥ 0.39). Calf weights and ADG did not differ through 108 DPP ( ≥ 0.77) or for the preweaning period ( ≥ 0.33). Plasma Se concentration was adequate for all cows and did not differ among treatments for cows ( ≥ 0.37) or calves ( ≥ 0.90). Liver Se concentrations in cows fed inorganic or organic Se were greater than in control cows ( < 0.01). Longissimus muscles biopsies taken from progeny at 108 DPP also did not differ between treatments ( = 0.45). In conclusion, dietary Se source did not affect cow performance, milk production, or reproductive efficiency. Organic Se decreased milk protein and increased milk lactose but did not alter preweaning performance of progeny from Se-adequate cows.

Intestinal gene expression profiles of piglets benefit from maternal supplementation with a yeast mannan-rich fraction during gestation and lactation.

The objective was to study the effect of maternal supplementation with a yeast cell wall-based product containing a mannan-rich fraction (MRF) during gestation and lactation on piglet intestinal gene expression. First parity sows were fed experimental gestation and lactation diets with or without MRF (900 mg/kg). After farrowing, piglets were fostered within treatment, as necessary. Sow and litter production performance data were collected until weaning. On day 10 post farrowing, jejunum samples from piglets were collected for gene expression analysis using the Affymetrix Porcine GeneChip array. Most performance parameters did not differ between the treatments. However, protein (P<0.01), total solids less fat (P<0.03) and the concentration of immunoglobulin G (IgG) in milk were greater (P<0.05) in the MRF-supplemented group. Gene expression results using hierarchical clustering revealed an overall dietary effect. Further analysis elucidated activation of pathways involved in tissue development, functioning and immunity, as well as greater cell proliferation and less migration of cells in the jejunum tissue. In conclusion, feeding the sow MRF during pregnancy and lactation was an effective nutritional strategy to bolster colostrum and milk IgG that are essential for development of piglet immune system and gut. In addition, the gene expression patterns affected by the passive immunity transfer showed indicators that could benefit animal performance long term.

Relationships between residual feed intake and hepatic mitochondrial function in growing beef cattle.

The objective of this study was to evaluate the relationship between hepatic mitochondrial function and residual feed intake (RFI) in growing beef cattle. In Trial 1, RFI was measured in 29 Angus heifers (initial BW = 258.0 ± 24.9 kg) from divergent IGF-I selection lines created at the Eastern Agricultural Research Station (The Ohio State University) fed a grain-based diet (calculated ME = 2.85 Mcal/kg DM). In Trial 2, RFI was measured in 119 Santa Gertrudis steers (initial BW = 308.4 ± 28.1 kg) fed a roughage-based diet (calculated ME = 2.21 Mcal/kg DM). At the end of the RFI measurement period, cattle in Trial 1 (n = 7 low RFI and n = 7 high RFI) and in Trial 2 (n = 6 low RFI and n = 8 high RFI) with measures of RFI exceeding 0.5 (Trial 1) or 1.0 (Trial 2) SD from the mean RFI were selected to measure mitochondrial function. Overall ADG, DMI, and RFI were 1.19 ± 0.15, 9.31 ± 1.12, and 0.00 ± 0.63 kg/d and 0.83 ± 0.16, 9.48 ± 1.00, and 0.00 ± 0.86 kg/d in Trial 1 and 2, respectively. Cattle with low RFI consumed 13 and 24% less (P < 0.05) DM and had 14 and 56% greater (P < 0.05) G:F than cattle with high RFI in Trial 1 and 2, respectively, even though ADG and BW were similar (P > 0.10). In Trial 1, cattle with low RFI tended (P = 0.06) to have greater state 3 respiration rates than cattle with high RFI, but state 3 respiration rates were similar (P > 0.10) between cattle with low and high RFI in Trial 2. In both trials, cattle with low RFI had greater (P < 0.05) acceptor control ratios than their high RFI counterparts. The respiratory control ratio tended (P = 0.09) to be greater for cattle with low RFI compared with high RFI cattle in Trial 1, but no difference (P > 0.10) was observed in Trial 2. Proton-leak kinetics were similar (P > 0.05) between cattle with low and high RFI in both trials. These data suggest that ADP has greater control of oxidative phosphorylation in liver mitochondrial of cattle with low RFI compared to their high RFI counterparts.

Recovery of insulin sensitivity in mature horses after a 3 week course of dexamethasone therapy.

REASONS FOR PERFORMING THE STUDY: Dexamethasone is an anti-inflammatory drug commonly used in equine medicine. Insulin sensitivity decreases with prolonged dexamethasone administration, but little information is available about the duration of this side effect after long-term treatment ends. OBJECTIVES: To determine how long it takes for blood glucose, insulin and markers of insulin sensitivity to return to normal ranges after extended dexamethasone treatment has ceased. STUDY DESIGN: Experimental study. METHODS: Eight healthy, mature, mixed-breed horses received 0.04 mg/kg bwt/day oral dexamethasone for 21 days. Blood samples were taken weekly during dexamethasone treatment (Days -21, -14 and -7). Following the final dose of dexamethasone on Day 0, blood samples were taken on Days 1-6, 8, 10, 12, 15 and 22. Day -21 represents baseline or normal blood predexamethasone. RESULTS: On Day 1, plasma glucose and insulin concentrations and the modified insulin-to-glucose ratio (a proxy for pancreatic ß cell responsiveness) were higher and the reciprocal of the square root of insulin (a proxy for the estimate of insulin sensitivity) was lower, in comparison with Day -21 values. Blood glucose concentrations dropped and returned to Day -21 values by Day 2. Insulin concentrations remained elevated until Day 3. Values for the modified insulin-to-glucose ratio decreased and returned to Day -21 concentrations by Day 4. Values for the reciprocal of the square root of insulin did not return to Day -21 values until Day 15. CONCLUSIONS: These results indicate that, in contrast to blood glucose concentrations, which return to normal quickly (within 2 days after treatment ends), the pancreatic insulin-secreting response has a delayed recovery.

Organic trace mineral levels in the first 96-h post-hatch impact growth performance and intestinal gene expression in broiler chicks.

Alterations in nutrient intake in the avian neonatal posthatch period can impact development, performance, and metabolism in adulthood. Very little is known about how mineral levels during the post-hatch period affect or "program" gene expression patterns later in life. The objective of this study was to determine the effect of post-hatch (0 to 96 h) dietary mineral supplementation on performance, tissue mineral content, and intestinal gene expression profiles in 21-day-old broiler chicks. One-day-old chicks were randomly assigned to one of two treatment groups consisting of N (organic Zn, Cu, and Mn provided at 100 % of recommendations (National Research Council 1994)) and/or L (organic Zn, Cu, and Mn provided at 20 % of recommendations (National Research Council 1994)) diets fed in two intervals (days 1­4, days 5­21) as follows: (1)N­Lor (2)L­L. Performance parameters did not differ between treatments except that body weight gain was greater (P < 0.05) in L­L birds than N­L birds over the experimental period. Bone mineral content was similar for both treatments at day 21. Intestinal gene expression profiling was examined using the Affymetrix GeneChip Chicken genome array. Ingenuity pathway analysis revealed differences in gene expression profiles between N and L treatments at day 5. At day 21, profiles were unique between N­L and L­L, suggesting that the diet fed until day 4 had an impact on gene expression patterns at day 21 even when birds were fed the same diets day 5­day 21. In this study, we demonstrated that diets fed for the 96 h post-hatch had long-term effects on gene expression, providing unique information as to why post-hatch diets are so important for the longterm bird health and productivity.

Comparison of gene expression profiles of the jejunum of broilers supplemented with a yeast cell wall-derived mannan oligosaccharide versus bacitractin methylene disalicylate.

1. The addition of yeast cell wall (YCW) mannan fractions or low concentrations of antibiotics to the diet of broilers positively affects gut health by improving intestinal cell morphology and improves feed efficiency and performance; however the exact mechanisms are unclear. Based on these production responses, the objective of this study was to compare the effects of supplementing YCW and bacitracin methylene disalicylate (BMD) in the diet on mRNA levels in the jejunum of 6-week-old broilers. 2. Dietary treatments were a maize-soya control diet and the control diet with the addition of YCW or BMD. Birds (n = 7) from each dietary treatment were randomly selected and killed at d 42. Whole jejunum (with serosa) samples were collected for RNA isolation. Gene expression analysis was performed using the AffymetrixGeneChip Chicken Genome Array (Santa Clara, CA, USA). 3. Supplementation with YCW resulted in 928 genes that were significantly changed (456 down-regulated, 472 up-regulated) and supplementation with BMD resulted in 857 genes that significantly changed (408 down-regulated, 449 up-regulated). In addition, 316 genes were significantly changed by both YCW and BMD (146 down-regulated, 170 up-regulated). 4. BMD increased the expression of genes involved in lipid and carbohydrate metabolism and decreased expression of genes associated with T-helper cell pathways. Gene expression profiles from birds fed on diets containing YCW showed changes on a genomic level that correspond to slower gut cell turnover and therefore increased energy preservation for growth. 5. In conclusion, supplementation with BMD or YCW had similar influences on the number of differentially expressed genes in the jejunum. Biological functions common to both YCW and BMD with positive activation scores included antiviral response and antimicrobial response. Genes that were affected by BMD or YCW classified into both different and common biological functions and pathways related to improved metabolism and health in the jejunum.

Antibodies to heteromeric glycolipid complexes in multifocal motor neuropathy.

BACKGROUND: Measurement of anti-GM1 IgM antibodies in multifocal motor neuropathy (MMN) sera is confounded by relatively low sensitivity that limits clinical usefulness. Combinatorial assay methods, in which antibodies react to heteromeric complexes of two or more glycolipids, are being increasingly applied to this area of diagnostic testing. METHODS: A newly developed combinatorial glycoarray able to identify antibodies to 45 different heteromeric glycolipid complexes and their 10 individual glycolipid components was applied to a randomly selected population of 33 MMN cases and 57 normal or disease controls. Comparison with an enzyme-linked immunosorbent assay (ELISA) was conducted for selected single glycolipids and their complexes. RESULTS: By ELISA, 22/33 MMN cases had detectable anti-GM1 IgM antibodies, whereas 19/33 MMN samples were positive for anti-GM1 antibodies by glycoarray. Analysis of variance (anova) revealed that of the 55 possible single glycolipids and their 1:1 complexes, antibodies to the GM1:galactocerebroside (GM1:GalC) complex were most significantly associated with MMN, returning 33/33 MMN samples as positive by glycoarray and 29/33 positive by ELISA. Regression analysis revealed a high correlation in absolute values between ELISA and glycoarray. Receiver operator characteristic analysis revealed insignificantly different diagnostic performance between the two methods. However, the glycoarray appeared to offer slightly improved sensitivity by identifying antibodies in four ELISA-negative samples. CONCLUSIONS: The use of combinatorial glycoarray or ELISA increased the diagnostic sensitivity of anti-glycolipid antibody testing in this cohort of MMN cases, without significantly affecting specificity, and may be a useful assay modification for routine clinical screening.

Effects of organic and inorganic dietary selenium supplementation on gene expression profiles in oviduct tissue from broiler-breeder hens.

Selenium (Se) is an essential component of at least 25 selenoproteins involved in a multitude of physiological functions, including reproduction. However, relatively little is known about the mechanisms by which Se exerts its physiological effects in reproductive tissue. The objective of this study was to compare the effect of long-term inorganic Se (sodium selenite, SS) and organic yeast-derived Se (Sel-Plex(®), SP) supplementations on tissue Se content and gene expression patterns in the oviduct of broiler-breeder hens. Hens were randomly assigned at 6 weeks of age to one of the three treatments: basal semi-purified diet (control), basal diet+0.3 ppm Se as SP or basal diet+0.3 ppm Se as SS. At 49 weeks, oviduct tissue from hens randomly selected from each treatment (n=7) was analyzed for Se content and gene expression profiles using the Affymetrix Chicken genome array. Gene expression data were evaluated using GeneSpring GX 10.0 (Silicon Genetics, Redwood, CA) and Ingenuity Pathways Analysis software (Ingenuity Systems, Redwood City, CA). Oviduct Se concentration was greater with Se supplementation compared with the control (P≤0.05) but did not differ between SS- and SP-supplemented groups. Gene expression analysis revealed that the quantity of gene transcripts associated with energy production and protein translation were greater in the oviduct with SP but not SS supplementation. Targets up-regulated by SP, but not SS, included genes encoding several subunits of the mitochondrial respiratory complexes, ubiquinone production and ribosomal subunits. SS hens showed a decrease in transcripts of genes involved in respiratory complexes, ATP synthesis and protein translation and metabolism in oviduct relative to control hens. In this study, although tissue Se concentrations did not differ between hens fed SS- and SP-supplemented diets, expression patterns of genes involved in energy production and protein synthesis pathways differed between treatments. These variations may partially explain the differences in reproductive performance reported in hens fed different forms of Se.

A comparative transcriptomic study of vitamin E and an algae-based antioxidant as antioxidative agents: investigation of replacing vitamin E with the algae-based antioxidant in broiler diets.

Previous study indicated that inclusion of an algae-based antioxidant as an antioxidative agent [EconomasE, Alltech, Nicholasville, KY; EcoE] significantly reduced the amount of vitamin E (VE) required in broiler diets without compromising performance and meat quality. To assess the mechanisms related to the VE-saving activity of EcoE, as well as other potential functions related to EcoE and VE supplementation, we analyzed gene expression profiles of breast muscle from broilers fed a control diet, the control diet + 50 IU of VE/kg, the control diet + 100 IU of VE/kg, or the control diet + 200 g of EcoE/ton. Evaluation of the serum antioxidant capacity indicated that dietary supplementation of either a high level of VE (50 or 100 IU of VE/kg) or EcoE significantly improved bird antioxidant status. Analysis of gene expression profiles indicated that expression of 542 genes of the breast muscle were altered (P < 0.05, fold change >1.2) by dietary treatments, of which a significant part were commonly regulated by EcoE and VE (especially the control diet + 50 IU of VE/kg). In addition to the process of cellular oxidation, gene ontology analysis indicated the involvement of EcoE and VE on cell morphology, skeletal and muscular system development and function, immune response, and multiple metabolic processes, including lipid, carbohydrate, and drug metabolism. Results of this experiment indicate that the biological roles of high VE, including its activity as an antioxidant, can be greatly mimicked at the transcriptional level by EcoE, and they suggest a relationship of functional redundancy between VE and EcoE in the broiler diets.

Students' experiences and perceptions of the use of portfolios in UK preregistration dietetic placements: a questionnaire-based study.

BACKGROUND: Reflective portfolios were introduced to dietetic practice placements in 2005, providing evidence for learning outcomes achieved and acting as a summative assessment tool. Portfolios may measure clinical competence more effectively than conventional examinations, but can be time consuming and subjective. The present study investigated current dietetic students' experiences and perceptions of the use and effectiveness of portfolio based learning and assessment during practice placements. METHODS: Current UK dietetic students, who had completed a B or B and C placement, were invited to participate, via University course leaders, in an online questionnaire exploring opinions and experiences of portfolio preparation, generation, assessment, and personal and professional development and reflection. RESULTS: One hundred and fourteen students from 11 Universities participated. Seventy-seven percent would have liked more information about portfolio construction prior to placement. Eighty percent of students believed that reflection helped monitor their strengths and weaknesses. Perceived reflective skills were significantly positively correlated with students' perceived writing skills (P < 0.0005) and academic ability (P = 0.002). Of the respondents, 92% agreed the portfolio was a valuable learning experience; however, 76% agreed that the amount of paperwork involved was excessive and 67% felt there were inconsistencies in portfolio assessment by different supervisors. CONCLUSIONS: Portfolio weaknesses identified are not specific to dietetics but are inherent to portfolio assessment across many professions. The introduction of national standardised assessment processes, practices, tools and training for assessors may help improve inter-departmental and inter-rater reliability, respectively.

Heteromeric glycolipid complexes as modulators of autoantibody and lectin binding.

Glycolipids act as receptors for a wide range of antibodies, lectins and microbes. It has long been recognised that the local topography of glycolipids in the plasma membrane is critical to these recognition events, although the biological basis for this has been relatively under-investigated. Within the last five years, emerging evidence indicates that hetero-dimeric clusters of different glycolipids can form highly distinct and specific epitopes for antibody and lectin binding. The initial observation that these ganglioside complexes (GSC) could either dramatically enhance or equally well inhibit the binding of neuropathy sera has now been reproduced for a number of other lectins, including siglecs and bacterial toxins. Here we review the initial discovery of GSC as antibody binding domains and the subsequent studies delineating their broader functional importance. Potential mechanisms underlying these effects are considered, although much remains to be investigated and explained. However, the implications for this field are potentially widespread, ranging from glycoarray design, structural biology and membrane biophysics, through to the biological consequences of glycolipid complex organisation in plasma membranes.

Body weight loss in beef cows: II. Increased antioxidant messenger ribonucleic acid levels in skeletal muscle but not erythrocyte antioxidant activity.

Twenty-six Angus-cross cows were used to examine the effect of BW loss (WL) on skeletal muscle and erythrocyte markers of oxidative stress. Serum NEFA concentrations, erythrocyte superoxide dismutase, and glutathione peroxidase activities were measured during WL and BW maintenance. Real-time reverse-transcription-PCR was used to determine mRNA levels of antioxidant genes during both periods to assess skeletal muscle response to WL. Body weight loss resulted in elevated serum NEFA concentrations but no change in erythrocyte superoxide dismutase and glutathione peroxidase activities. During WL, mRNA levels of the antioxidant genes glutathione peroxidase 4, mitochondrial superoxide dismutase, thioredoxin reductase 1, and selenoprotein W increased. Abundance of mRNA of genes involved in antioxidant signaling, specifically, PPARgamma coactivator-1 alpha, nuclear respiratory factor 1, estrogen-related receptor alpha, and tumor protein 53, was also increased. In summary, during WL cows had no change in peripheral antioxidant enzyme activity, but mRNA abundance of proteins involved in protecting the body from oxidative stress increased in skeletal muscle. During times when NEFA are used as a fuel source, signals such as mild reactive oxygen species production or increased concentration of lipid by-products activate the transcription of nuclear signaling molecules such as PPARgamma gamma coactivator-1 alpha, nuclear respiratory factor 1, estrogen-related receptor alpha, and tumor protein 53. These genes work to activate antioxidant genes such as mitochondrial superoxide dismutase, glutathione peroxidase 4, and thioredoxin reductase 1 to aid in the detoxification of reactive oxygen species. These data suggest an important role for antioxidant genes to protect cattle that are mobilizing body fat.

Body weight loss in beef cows: I. The effect of increased beta-oxidation on messenger ribonucleic acid levels of uncoupling proteins two and three and peroxisome proliferator-activated receptor in skeletal muscle.

Twenty-six Angus-cross cows were studied during BW loss (WL) and BW maintenance (WM) to examine the effects of elevated beta-oxidation on mRNA levels of NEFA-responsive signaling molecules in skeletal muscle. At the end of the WL and WM sampling periods, muscle biopsies were removed from the biceps femoris and mRNA levels were measured using real-time PCR. In comparison with WM, cows undergoing WL had elevated mRNA levels of carnitine palmitoyltransferase 1 (4.6-fold), fatty acid binding protein 3 (2.0-fold), and acyl-coenzyme A oxidase 1 (2.8-fold), all of which are indicators of beta-oxidation. Levels of mRNA of the NEFA-responsive signaling molecules PPAR alpha, delta, and gamma increased 2.0-fold, 2.2-fold, and 1.84-fold, respectively, during WL. Uncoupling proteins 2 and 3 also had increased mRNA (3.0-fold and 6.0-fold, respectively) during WL, but Western blot analysis found no changes in protein abundance of uncoupling protein 3. Uncoupling protein expression can be directly stimulated by elevated NEFA, potentially to protect cells from damage by lipid oxidation by-products. Thus, an increase in mRNA levels of genes involved in beta-oxidation of fatty acids and fatty acid by-products occurs during BW loss in beef cattle. These data support previous findings in nonruminants and suggest that these genes play a role in the same physiological processes in ruminants.

Expression of the hypothalamic transcription factor Nhlh2 is dependent on energy availability.

Mice with a deletion of the hypothalamic basic helix-loop-helix transcription factor Nhlh2 display adult onset obesity, implicating Nhlh2 in the neuronal circuits regulating energy availability. Nhlh2 colocalises with the hypothalamic thyrotrophin-releasing hormone (TRH) neurones in the paraventricular nucleus (PVN) and pro-opiomelanocortin (POMC) neurones in the arcuate nucleus. We show that Nhlh2 expression is significantly reduced in response to 24-h food deprivation in the arcuate nucleus, PVN, lateral hypothalamus, ventromedial hypothalamus (VMH) and dorsomedial hypothalamus (DMH). Food intake for 2 h following deprivation stimulates Nhlh2 expression in the arcuate nucleus and the PVN, and leptin injection following deprivation results in increased Nhlh2 expression in the arcuate nucleus, PVN, lateral hypothalamus, VMH, and DMH. Hypothalamic Nhlh2 expression in response to leptin injection is maximal by 2 h. Following leptin injection, Nhlh2 mRNA colocalises in POMC neurones in the arcuate nucleus and TRH neurones in the PVN. Nhlh2 mRNA expression in POMC neurones in the arcuate nucleus and TRH neurones in the PVN is reduced with energy deprivation and is stimulated with food intake and leptin injection. Modulation of POMC expression in response to changes in energy availability is not affected in mice with a targeted deletion of Nhlh2. However, deletion of Nhlh2 does result in loss of normal TRH mRNA expression in mice exposed to food deprivation and leptin stimulation. These data implicate Nhlh2 as a regulatory target of the leptin-mediated energy availability network of the hypothalamus, and TRH as a putative downstream target of Nhlh2.

Genetic analysis of NHLH2 and its putative role in bovine body weight control.

The control of energy balance is fundamental to adult animals and is necessary for weight gain/loss, reproductive capacity and general health. In mice, targeted deletion of the neuronal transcription factor Nhlh2 results in adult-onset obesity because of reduced exercise and infertility because of reduced sexual behaviour. Nhlh2 (NHLH2 for humans) is expressed in the hypothalamus, particularly in neurons that have been shown to regulate energy balance. We have cloned the bovine Nhlh2 gene (bNHLH2) and we have shown that bNHLH2 is also expressed in the hypothalamus. Phylogenetic analysis of Nhlh2 reveals that it is very highly conserved in humans, mice, chimps and cattle, and found in organisms with simpler nervous systems, including Caenorhabditis elegans and Drosophila. Using a cattle-human comparative map and online databases, we have evidence that bNHLH2 is located near a quantitative trait locus for marbling on bovine chromosome 3 between microsatellite markers BM723 and BMS963. Cloning of the bNHLH2 gene from Holstein cattle and a mixed breed individual and comparison with Hereford sequences shows that the gene is highly conserved among bovine breeds.

Building a vision for the future: strategic planning in a shared governance nursing organization.

Today's health care delivery environment is marked by extreme turbulence and ever-increasing complexity. Now, more than ever, an organization's strategic plan must do more than outline a business plan. Rather, the strategic plan is a fundamental tool for building and sustaining an organizational vision for the future. The strong, dynamic strategic plan (1) represents a long-range vision for improving organizational performance, (2) provides a model for planning and implementing structures and processes for the management of outcomes, (3) reflects and shapes the organizational culture and customer focus, (4) provides decision support for difficult operational choices made day to day, and (5) integrates and aligns the work of the organization. This article describes the development, implementation, and evaluation of a methodology for strategic planning within a shared governance nursing organization. Built upon the strategic plan of the hospital, the process undertaken by the nursing organization reflects the following commitments: (1) to develop a strategic plan that is meaningful and part of daily work life at all levels of the nursing organization, (2) to make the plan practical and realistic through incremental building, (3) to locate and articulate accountability for each step, and (4) to build in a process for checking progress toward goal achievement and readjusting the plan as necessary.

Temperature determination in the frozen region during cryosurgery of rabbit liver using MR image analysis.

Cryosurgery currently is being used clinically to treat tumors in internal organs such as the liver and prostate. Although performed at present under ultrasound monitoring, magnetic resonance imaging (MRI)-guidance of these procedures not only permits monitoring of the frozen region during cryosurgery but also makes it possible to determine the temperature distribution in the frozen region, which is not possible using ultrasound monitoring. A good estimate of the region of destruction in the tissue can be obtained from correlating the temperature distribution and the time course of the freezing with the image of the frozen region. Unfortunately, MR pulse sequence-based temperature determination techniques such as diffusion, relaxation time, and chemical shift cannot be used for measuring the temperature in the frozen region because the T2 of the frozen regions is so short that there is effectively no RF signal from the frozen region. This paper describes a numerical technique for determining the two dimensional temperature distribution in the frozen region during MR image-guided cryosurgery of normal liver in rabbits. The technique involves solving the energy equation numerically in the frozen region to determine the temperature distribution there. The boundary conditions needed to solve the equation are determined from MR images of the frozen tissue during cryosurgery and from the measured temperature of the cryoprobe. The calculated temperature in the frozen region is then correlated with the damaged region (cryolesion) determined from post mortem histologic evaluation.