Reference gene selection for reverse transcription quantitative polymerase chain reaction in chicken hypothalamus under different feeding status.

This study was designed to investigate the stability of 10 candidate reference genes, namely ACTB, B2M, GAPDH, HMBS, LBR, POLR2B, RN18S, RPS17, TBP, and YWHAZ for the normalization of gene expression data obtained by quantitative real-time polymerase chain reaction (qPCR) in studies related to feed intake of chicken. Samples were isolated from hypothalamus under three different nutritional status (ad libitum, fasted for 24 hr, fasted for 24 hr then refed for 2 hr). Five different algorithms were applied for the analysis of reference gene stability: BestKeeper, geNorm, NormFinder, the comparative ΔCt method, and a novel approach using multivariate linear mixed-effects modelling for stable reference gene selection. TBP and POLR2B were identified as the two most suitable and B2M and RN18S as the two least stable reference genes for normalization. Despite our review, the current literature showing that RN18S is one of the most commonly used reference gene in chicken gene expression studies, its applicability for normalization should be evaluated before each qPCR experiment.

Effect of nano-sized, elemental selenium supplement on the proteome of chicken liver.

The nano-sized (100-500 nm) selenium has higher bioavailability and relatively lower toxicity compared to other selenium forms. The objective of the present study was to compare liver proteome profiles of broiler chicken fed with control diet without Se supplementation and diet supplemented with nano-Se with 4.25 mg/kg DM. Differential proteome analyses were performed by two-dimensional gel electrophoresis (2D-PAGE) followed by tryptic digestion and protein identification by liquid chromatography-mass spectrometry (LC-MS). Seven hundred and eight spots were detected, and 18 protein spots showed significant difference in their intensity (p < 0.05) between the two groups. In response to nano-Se supplementation, the expression of 8 proteins was higher, and 5 proteins were lower in nano-Se supplemented group compared to control group. The functions of the differentially expressed proteins indicate that the high dose of selenium supplementation induced a dietary stress. Selenium supplementation may influence the metabolism of fatty acids and carbohydrates and antioxidant system, and increase the quantity of cytoskeletal actin and the expression of actin regulatory protein as well.

Reproduction Efficiency and Health Traits in Dorper, White Dorper, and Tsigai Sheep Breeds under Temperate European Conditions.

The objective of the current pilot study was to evaluate the reproductive performance and health indicators in Dorper, White Dorper, and Tsigai breed ewes managed semi-intensively under European temperate conditions. A total of 544 ewe-year units were observed, with ewes (ranging from 1.5 to 8 years of age) managed under identical rearing conditions for a period of two consecutive production cycles (2012 through 2013 and 2013 through 2014). In general, significant (p≤0.001) genotype-related disparities were found in occurrence rates for all health parameters taken into study. Clinical mastitis incidence was significantly lower (p≤0.05) in Dorper (9.4%) and White Dorper (10.8%) breeds compared to that of Tsigai ewes (17.4%). Significant differences (p≤0.05) for lameness were found between Dorper and Tsigai breeds, with occurrence rates of 8.0% and 2.9%, respectively. Incidence for pneumonia and abortion was not influenced (p>0.05) by the ewes' genotype. Litter size was significantly lower (p≤0.05) in White Dorper breed than for Dorper and Tsigai ewes, of 1.21, 1.40, and 1.45, respectively. Conception rates and lambs survival were not affected (p>0.05) by genotype. Results suggest that South African Dorper and White Dorper sheep breeds have adapted well to the specific rearing conditions.

Genistein induces phosphorylation of cAMP response element-binding protein in neonatal hypothalamus in vivo.

We investigated the effect of the phytoestrogen, genistein and 17beta-oestradiol on cAMP response element-binding protein (CREB) phosphorylation in the neonatal female rat hypothalamus in vivo using western blot analysis and immunohistochemistry. Although CREB expression was insensitive to the compounds we tested, administration of genistein and 17beta-oestradiol induced rapid CREB phosphorylation (< 15 min) in the hypothalamus and its level remained elevated at 4 h. Quantitative immunohistochemical analysis showed that genistein and 17beta-oestradiol had no effect on CREB phosphorylation in the magnocellular subdivision of paraventricular nucleus. By contrast, genistein induced a dose-dependent increase in CREB phosphorylation in the medial preoptic area (mPOA) and anteroventral periventricular nucleus (AVPV). Administration of 17beta-oestradiol also caused a rapid, dose-dependent increase in CREB phosphorylation in the hypothalamus, mPOA and AVPV. These results demonstrate that genistein induces oestrogen-like rapid action on CREB phosphorylation in the neonatal central nervous system in vivo.

Searching for DNA markers for milk production and composition on chromosome 6 in sheep.

Several milk protein polymorphisms are potential tools for selection in dairy ruminants. However, research results for dairy sheep are not as conclusive as those for goats or cattle and are often controversial. The main objective of this study was to find and later use molecular genetic markers in selection to improve milk production and milk composition in Awassi ewes. Chromosome 6 was chosen because several studies have reported the presence of significant quantitative trait loci (QTL) affecting milk production traits on ovine and bovine chromosome 6. Altogether, genotypes for 13 microsatellite loci were determined for 258 ewes, which were purebred Awassi or Awassi-Merino crosses. Phenotypic data were lactation yield of milk, milk fat, protein and lactose (kg), average milk protein and fat percentage and average somatic cell count. Five out of the 13 microsatellites showed significant association with at least one of the examined traits.

Changes in MPF and MAPK activities in porcine oocytes activated by different methods.

The effect of different oocyte activation methods on the dynamics of M-phase promoting factor (MPF) and mitogen-activated protein kinase (MAPK) activity in porcine oocytes were examined. Three activativation methods were tested: (1) electroporation (EP); (2) electroporation combined with butyrolactone I (BL), an inhibitor of cdc2 and cdk2 kinases; (3) electroporation followed by a treatment with cycloheximide (CHX), a protein synthesis blocker. The activity of cdc2 in MII oocytes was 0.067+/-0.011pmol/oocyte/min (mean+/-S.E.M.), which by 1h decreased in every treatment group (P<0.05) and stayed at low levels until 6h post-activation, approximately the time of pronuclear formation. The initial MAPK activity (0.123+/-0.017pmol/oocyte/min) also decreased 1h after each type of activation treatment (P<0.005). However, in the electroporation only group, activity reached its lowest level at 3h; thereafter, it started to recover and at later time points, MAPK activity did not differ from that in non-treated oocytes (P>0.1). In contrast, oocytes where electroporation was followed by protein kinase or protein synthesis inhibition had low MAPK activity by the time pronuclei were to be formed. Pronuclear formation in these groups (86.3+/-3.3% for EP+BL and 87.6+/-3.7% for EP+CHX) was higher compared to that found in the EP-only oocytes (69.4+/-3.3%; P<0.05). These findings demonstrated that electroporation alone efficiently triggered the inactivation of MPF but not that of MAPK. In order to achieve low MAPK activity to allow high frequency of pronuclear formation, electroporation should be followed by a treatment that inhibits protein synthesis or specific protein kinases. The combined activation methods provided stimuli that efficiently induced both MPF and MAPK inactivation and triggered pronuclear formation with high frequencies.

Investigation of two candidate genes for meat quality traits in a quantitative trait locus region on SSC6: the porcine short heterodimer partner and heart fatty acid binding protein genes.

A highly significant quantitative trait locus (QTL) on pig chromosome 6, affecting intramuscular fat (IMF), has previously been detected by our group and others. Two genes of positional and biological interest, the small heterodimer partner (SHP; NR0B2) and the heart fatty acid binding protein (FABP3; H-FABP), were investigated for meat quality traits and IMF respectively. SHP was partially sequenced (GenBank: DQ002896 and DQ002897) and mapped to the QTL region on porcine chromosome 6, affecting IMF. The map shows no recombination between SHP and FABP3, which was previously mapped to the same QTL region. Twelve single nucleotide polymorphisms were detected in the sequenced region of SHP gene. Haplotype information was used to investigate association between genetic variation and different meat quality traits. SHP haplotype combinations were found to have significant effect on connective tissue. However, further studies are needed to evaluate this possible association more effectively. The FABP3 is involved in fatty acid transport and has been studied as a candidate gene for IMF by several research groups. In our study, FABP3 genotypes were confirmed to be significantly associated with IMF in pigs. The average content of IMF in our population was 1.6%, which may indicate that the FABP3 polymorphism explains as much as 30-35% of the variation in IMF in our pig cross-population.

[Reliability and use of prostate-specific antigen in the management of patients with prostatic cancer]. / A prostata specificus antigen megbízhatósága és haszna a prostata carcinomás betegek gondozásában.

The authors are reporting the clinical importance of prostate specific antigen. They concluded that there are false positive and false negative cases. The prostate specific antigen is high in prostatic hyperplasia therefore the distinction between the two disease is impossible on basis of the prostate specific antigen. Prostate specific antigen shows the metastatic cases better [correction of worse] than prostatic acid phosphatase. But prostate specific antigen detects the changes of the clinical course of the disease well and shows the progression sooner. The authors have concluded that prostate specific antigen can not replace phosphatases and bone-scanning in the diagnosis and follow up of patients with prostate carcinoma.

Different cholesterol fractions, LCAT activity and lipid peroxides in the serum of children whose parents had early coronary heart disease.

Children whose parents had early coronary heart disease were investigated for lipid abnormalities. In order to assess high risk parameters and the efficacy of the applied care, serum total cholesterol (TC), total triglyceride (TT), total free cholesterol (FC) high density lipoprotein cholesterol (HDLC), high density lipoprotein free cholesterol (HDLFC), lipid peroxide (LP) levels, and lecithin: cholesterol acyl transferase (LCAT) activity were measured. Compared to a group of control children, the offspring of high risk subjects had increased TC, FC, and LP levels and decreased HDLC levels. After one year of preventative care all parameters normalized except the high FC level and elevated LCAT activity. The measurement of serum FC and LP levels seems to be a useful method for the determination of true high risk. The LCAT activity may show the efficacy of the dietary treatment.

Serum lipids, lipid peroxides and the care of children with high risk atherosclerotic family history.

Children of parents who had their first acute myocardial infarction (AMI) before the age of 45 years were investigated and divided into 2 groups. Group 1 consisted of 12 children: both their parents or one parent and grandparents suffered from AMI. Group 2 consisted of 55 children, each of whom had one parent with AMI. Serum levels of total cholesterol (TC) and lipid peroxide (LP) were higher in these children and HDL-cholesterol values were lower than those of 39 control children. The extent of serum TC and HDL-C differences was similar in both high risk groups but the serum LP level was significantly higher in group 1 than in group 2. The serum free cholesterol (FC) level was higher in group 1 than in controls. Children were treated according to a program giving advice on diet, physical exercise and health education for 1 year. At the end of this period the serum HDL-C level increased in both groups, while LP and TC levels decreased. The high serum FC level did not change.

In vitro study of local solubility of uric acid stones with lithium carbonate.

The possibilities of local dissolution of uric acid calculi with Na hydrocarbonate, K carbonate and lithium carbonate were examined in vitro. The most promising results were obtained with lithium carbonate. The necessity for studies of the compound in laboratory animals before its use in humans is emphasized.

Pyelolithotomy by the use of a hydrophilous macromolecular binding material ("pyeloform" pyelotomy).

The use of a synthetic macromolecular hydrophilous binding material for the extraction of multiple renal calculi (" Pyeloform " pyelotomy) is reported. An account of the results of animal studies in vitro is given and the possibilities of the method in are pointed out.

Applicability of biosuppressin as an urease-inhibitor.

Prevention of the formation of struvite and carbonate-apatite calculi, which rapidly increase in size and tend to recur, is of prime importance. One of the urease inhibitors, hydroxycarbamide, was studied in vitro. The results are favourable and justify further studies aimed at the local application of the inhibitor.