Sleeping Beauty-mediated knockdown of sheep myostatin by RNA interference

Myostatin is a negative regulator of skeletal muscle growth. Myostatin dysfunction therefore offers a strategy for promoting animal muscle growth in livestock production. Knockdown of myostatin was achieved by combining RNA interference and the Sleeping Beauty (SB) transposon system in sheep cells. Four targeting sites of sheep myostatin were designed and measured for myostatin silencing in sheep fetal fibroblasts by real-time PCR. The sh3 construct induced significant decrease of myostatin gene expression by 90% (P < 0.05). Myostatin silencing induced by SB-mediated sh3 was further tested in stably transfected cells. SB transposition increased the integration frequency of genes into sheep genomes and mediated a more efficient myostatin knockdown than random integration of sh3. We suggest that SB-mediated shRNA provides a novel potential tool for gene knockdown in the donor cells of animal cloning.     

Ice-free cryopreservation of heart valve allografts: better extracellular matrix preservation in vivo and preclinical results

The purpose of this study was evaluation of an ice-free cryopreservation method for heart valves in an allogeneic juvenile pulmonary sheep implant model and comparison with traditionally frozen cryopreserved valves. Hearts of 15 crossbred Whiteface sheep were procured in Minnesota. The valves were processed in South Carolina and the pulmonary valves implanted orthotopically in 12 black faced Heidschnucke sheep in Germany. The ice-free cryopreserved valves were cryopreserved in 12.6?mol/l cryoprotectant (4.65, 4.65, and 3.31?mol/l of dimethylsulfoxide, formamide and 1,2-propanediol) and stored at ?80°C. Frozen valves were cryopreserved by controlled slow rate freezing in 1.4?mol/l dimethylsulfoxide and stored in vapor-phase nitrogen. Aortic valve tissues were used to evaluate the impact of preservation without implantation. Multiphoton microscopy revealed reduced but not significantly damaged extracellular matrix before implantation in frozen valves compared with ice-free tissues. Viability assessment revealed significantly less metabolic activity in the ice-free valve leaflets and artery samples compared with frozen tissues (P?<?0.05). After 3 and 6?months in vivo valve function was determined by two-dimensional echo-Doppler and at 7?months the valves were explanted. Severe valvular stenosis with right heart failure was observed in recipients of frozen valves, the echo data revealed increased velocity and pressure gradients compared to ice-free valve recipients (P?=?0.0403, P?=?0.0591). Histo-pathology showed significantly thickened leaflets in the frozen valves (P?<?0.05) and infiltrating CD3+ T-cells (P?<?0.05) compared with ice-free valve leaflets. Multiphoton microscopy at explant revealed reduced inducible autofluorescence and extracellular matrix damage in the frozen explants and well preserved structures in the ice-free explant leaflets. In conclusion, ice-free cryopreservation of heart valve transplants at ?80°C avoids ice formation, tissue-glass cracking and preserves extracellular matrix integrity resulting in minimal inflammation and improved hemodynamics in allogeneic juvenile sheep.     

Polymorphism of 5′ regulatory region of ovine FSHR gene and its association with litter size in Small Tail Han sheep

Single nucleotide polymorphisms of 5?? regulatory region of follicle-stimulating hormone receptor (FSHR) gene were detected in two high prolificacy sheep breeds (Small Tail Han and Hu sheep) and two low prolificacy sheep breeds (Corriedale and Chinese Merino sheep) by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). The results indicated that there were three genotypes (AA, AB and BB) detected by primer 1 in Hu sheep while only one genotype (AA) in other three sheep breeds, and frequencies of AA, AB and BB genotypes in Hu sheep were 0.700, 0.225 and 0.075, respectively. There were three genotypes (EE, EF and EG) detected by primer 3 in Small Tail Han sheep while only EE genotype occurred in other three sheep breeds, and frequencies of EE, EF and EG genotypes in Small Tail Han sheep were 0.775, 0.200 and 0.025, respectively. No polymorphism was detected in four sheep breeds by primer 2 and primer 4. The sequencing results showed that there were two nucleotide mutations (g. ?681T>C and g. ?629C>T) in genotype BB compared with AA for primer 1. As for primer 3, two mutations (g. ?197G>A and g. ?98T>C) in genotype EF compared with EE and two mutations (g. ?200G>A and g. ?197G>A) in genotype EG compared with EE. The heterozygous ewes with EG or EF had 0.89 (P?<?0.05) or 0.42 (P?<?0.05) lambs more than homozygous ewes (EE genotype) in Small Tail Han sheep, respectively, while there was no significant difference on litter size between EG and EF ewes.     

Effects of dietary selenium and vitamin E on immune response and biological blood parameters of broilers reared under thermoneutral or heat stress conditions

A study was conducted using 360 broiler chickens to evaluate the effects of dietary vitamin E (0, 125 and 250 mg/kg), selenium (Se, 0, 0.5 and 1 mg/kg), or their different combinations on immune response and blood biological parameters of broilers raised under either thermoneutral (TN, 23.9 °C constant) or heat stress (HS, 23.9 to 37 °C cycling) conditions. Humoral immunity was assessed by intravenous injection of 7 % sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Heterophil to lymphocyte (H/L) ratio also determined as an indicator of stress. Furthermore, at the end of the experiment, birds were bled for determination of some biological parameters. There was a significant reduction in body weight and feed intake, but the feed conversion ratio increased when the birds were exposed to HS (P?<?0.05). Body weight and feed intake were not influenced significantly by dietary vitamin E and Se (P?>?0.05), whereas feed conversion was improved significantly by 125 mg/kg vitamin E (P?<?0.05). The liver and lymphoid organ weights as well as IgM and IgG, antibody titers for primary and secondary antibody responses to SRBC were reduced significantly under HS (P?<?0.05). Heat stress also resulted in a significant increase in H/L ratio (P?<?0.05). Dietary vitamin E resulted in improvement of primary and secondary antibody responses both in TN and HS broilers (P?<?0.05). The HS birds also showed an improved antibody titer in secondary response with high concentration of Se (P?<?0.05). Vitamin E and Se had interactive effects on anti-SRBC titers; however, no consistent differences were found between dietary levels during the study. The H/L ratio decreased by feeding vitamin E at both levels either under HS or TN conditions (P?<?0.05). The serum concentrations of glucose, triglycerides, total cholesterol, and LDL-cholesterol were increased but serum HDL-cholesterol decreased in HS broilers (P?<?0.05).     

Polymorphisms of KiSS-1 and GPR54 genes and their relationships with litter size in sheep

The KiSS-1 and GPR54 genes were studied as candidate genes for the prolificacy in sheep. Four pairs of primers were designed to detect single nucleotide polymorphisms of exon 1 of KiSS-1 gene and exon 1, exon 2 and partial exon 5 of GPR54 gene in high fecundity breeds (Small Tail Han and Hu sheep) and low fecundity breeds (Dorset, Texel and Corriedale sheep) by PCR-SSCP. Polymorphisms in exon 1 of KiSS-1 gene were detected in prolific Small Tail Han sheep (AA, AB and BB genotypes) and Hu sheep (AA and CC genotypes), no polymorphism was found in low fecundity sheep breeds (only AA genotype). Polymorphisms in exon 2 of GPR54 gene were detected in prolific Hu sheep (DD and EE genotypes) and no polymorphism was found in prolific Small Tail Han sheep and low fecundity sheep breeds (only DD genotype). No polymorphism was detected in exon 1 and partial exon 5 of GPR54 gene in five sheep breeds. The polymorphic genotypes were sequenced. While compared the BB genotype with the AA genotype, one nucleotide mutation (G1035A) was detected, which resulted in amino acid change, Val25Met. Five nucleotide mutations were detected from AA to CC genotype (C981T, C996T, T997C, C1034G, C1039T), and among them four caused amino acid changes, that is, Arg7Trp, Phe12Leu, Asn24Lys, Ala26Val. While compared the EE genotype with the DD genotype, two nucleotide mutations (T2360C, A2411C) were detected, which gave rise to amino acid changes, Met90Thr and Asp107Ala, respectively. Genotype frequencies of AA, BB and AB were 0.62, 0.05 and 0.33 in Small Tail Han sheep, respectively. The Small Tail Han sheep ewes with genotype BB or AB had 0.88 (P?<?0.05) or 0.51 (P?<?0.05) lambs more than those with genotype AA; the Small Tail Han sheep ewes with genotype BB had 0.37 (P?>?0.05) lambs more than those with genotype AB. These results preliminarily indicated that the KiSS-1 gene may have some association with prolificacy in sheep.     

Effects of Selenium Supplementation on Expression of Endothelin-1 and Its Receptors in Pulmonary Microvascular Endothelial Cells from Chick Embryos

The objective of this study was to evaluate the effects of supplemental selenium (Se) on expression of endothelin-1 (ET-1) and its receptors in cultured chick embryos pulmonary microvascular endothelial cells (PMVECs). To accomplish this, PMVECs were treated in Se-deficient or Se-supplement (12, 24, 50, 100?ng/ml) culture medium for 48?h. Low Se medium was achieved by reducing serum concentrations and the essential growth factors were added. After the incubation, the effects of supplemental Se on ET-1 and its receptors gene expression were assessed by quantitative real-time PCR (qRT-PCR). Compared with the control group, our results showed that among the different concentrations of Se supplement, the levels of ET-1 gene expression treated with both the moderate Se doses (24, 50?ng/ml, P?<?0.01, P?<?0.01, respectively) and the high doses (100?ng/ml, P?<?0.05) were noticeably decreased, the low-dose group (12?ng/ml), which showed no changes. Meanwhile, Se supplement (24, 50, 100?ng/ml) was found to be effective in reducing the expression levels of ETA (P?<?0.01, P?<?0.05, P?<?0.05, respectively) in cultured PMVECs grown in low Se medium. However, there were no significant changes (P?>?0.05) in ETB mRNA levels during the cell proliferation. These observations indicated that Se may play both direct and indirect role in the regulation of ET-1 and its receptors gene expression and their production in avian PMVECs. Se supplement decreases in ET-1 and ETA production in Se-deficient PMVECs may partly explain the mechanism of the protective effects of the Se on the cardiovascular system.     

The effect of hyperammonemia on myostatin and myogenic regulatory factor gene expression in broiler embryos

Myogenesis is facilitated by four myogenic regulatory factors and is significantly inhibited by myostatin. The objective of the current study was to examine embryonic gene regulation of myostatin/myogenic regulatory factors, and subsequent manipulations of protein synthesis, in broiler embryos under induced hyperammonemia. Broiler eggs were injected with ammonium acetate solution four times over 48 h beginning on either embryonic day (ED) 15 or 17. Serum ammonia concentration was significantly higher (P<0.05) in ammonium acetate injected embryos for both ED17 and ED19 collected samples when compared with sham-injected controls. Expression of mRNA, extracted from pectoralis major of experimental and control embryos, was measured using real-time quantitative PCR for myostatin, myogenic regulatory factors myogenic factor 5, myogenic determination factor 1, myogenin, myogenic regulatory factor 4 and paired box 7. A significantly lower (P<0.01) myostatin expression was accompanied by a higher serum ammonia concentration in both ED17 and ED19 collected samples. Myogenic factor 5 expression was higher (P<0.05) in ED17 collected samples administered ammonium acetate. In both ED17 and ED19 collected samples, myogenic regulatory factor 4 was lower (P⩽0.05) in ammonium acetate injected embryos. No significant difference was seen in myogenic determination factor 1, myogenin or paired box 7 expression between treatment groups for either age of sample collection. In addition, there was no significant difference in BrdU staining of histological samples taken from treated and control embryos. Myostatin protein levels were evaluated by Western blot analysis, and also showed lower myostatin expression (P<0.05). Overall, it appears possible to inhibit myostatin expression through hyperammonemia, which is expected to have a positive effect on embryonic myogenesis and postnatal muscle growth.     

Litterfall production, decomposition and nutrient use efficiency varies with tropical forest types in Xishuangbanna, SW China: a 10-year study

Litterfall production, decomposition and nutrient use efficiency in three different tropical forest ecosystems in SW China were studied for 10 years. Annual mean litterfall production in tropical seasonal forest (TSF) (9.47?±?1.65 Mg ha^?1) was similar to that in man-made tropical forest (MTF) (9.23?±?1.29 Mg ha^?1) (P?>?0.05) but both were significantly lower than that in secondary tropical forest (STF) (12.96?±?1.71 Mg ha^?1) (P?<?0.05). The annual variation of litterfall was greater in TSF (17.4%, P?<?0.05) than in MTF (14.0%) or STF (13.2%). The annual mean decomposition rate of litterfall increased followed the order of MTF (2.72)?<?TSF (3.15)?<?STF (3.50) (P?<?0.05), which was not correlated with annual precipitation or annual mean temperature, but was rather related to litter quality. The nutrient use efficiency was found to be element-dependent and to vary significantly among the three forest types (P?<?0.05). These results indicate that litterfall production and decomposition rates in different tropical forest systems are related to plant species composition and are influenced strongly by coexisting species and their life stage (age) but less so by the species richness. Constructing multi-species and multistory man-made tropical forest is an effective way to enhance biological productivity and maintain soil nutrients on degraded tropical land.     

Combined Impact of Exercise and Temperature in Learning and Memory Performance of Fluoride Toxicated Rats

In previous studies, we investigated a link between high fluoride exposure and functional IQ deficits in rats. This study is an extension conducted to explore the combined influence of physical exercise and temperature stress on the learning ability and memory in rats and to assess whether any positive modulation could be attenuated due to exercise regimen subjected to F-toxicated animals at different temperatures. Accumulation of ingested fluoride resulted significant inhibition in acetylcholinesterase activity (P?<?0.05), plasma cortisol levels (P?<?0.05), and impaired the acquisition, performance, latency time, and retention in fluoride-exposed animals. Fluoride-toxicated rats took more number of sessions during the learning phase [F _{5, 35}?=?19.065; P?<?0.05] and post hoc analysis on the number of correct choices revealed that there was a significant effect of treatments [F _{5, 30}?=?15.763; P?<?0.05]; sessions [F _{8, 240}?=?58.698; P?<?0.05]; and also significant difference in the interactions [F _{40, 240}?=?1.583; P?<?0.05]. The latency data also revealed a significant difference between groups [F _{5, 30}?=?28.085; P?<?0.05]; time?=?[F _{8, 240}?=?136.314; P?<?0.05]; and there was a significant difference in the interactions [F _{40, 240}?=?2.090; P?<?0.05]. In order to ascertain if interdependence between fluoride concentrations and the foregoing free radical parameters, respective correlation coefficients were calculated and results clearly emphasize the positive role of exercise in the promotion of cognitive functions by decreasing fluoride levels in rat hippocampus. A significant recovery in cognitive function was noticed in all the exercised animals due to reduced burden of brain oxidative stress. In comparison to exercise regimens performed at different temperatures, high (35?°C) and low temperatures (20?°C) led to a slower acquisition and poor retention of the task when compared to thermo neutral temperatures (25 and 30?°C). Thus exercise up-regulate antioxidant defenses and promote learning abilities in fluorotic population.     

Effectiveness of Maifanite in Reducing the Detrimental Effects of Cadmium on Growth Performance, Cadmium Residue, Hematological Parameters, Serum Biochemistry, and the Activities of Antioxidant Enzymes in Pigs

This study was conducted to investigate the toxicity of cadmium and to evaluate the effectiveness of maifanite in preventing cadmium-induced adverse effects. Thirty-two crossbred pigs (Duroc × Landrace × Large white, sex balanced, 17.25?±?0.07 kg average body weight) were randomly allotted to one of four dietary treatments in a 2?×?2 factorial arrangement, with eight replicates per treatment and one pig per replicate. The dietary treatments included two cadmium (as CdCl₂) doses (0.32 and 30.49 mg/kg) and two maifanite doses (0 and 1 %). The results showed that pigs treated with cadmium decreased their average daily feed intake (P?<?0.05) and increased (P?<?0.05) the feed/gain ratio. Cadmium was found in the tissues of pigs that were fed with cadmium-contaminated diets, but the level of cadmium was much lower when maifanite was added to the cadmium-contaminated diets. Ingestion of diets that were artificially contaminated with cadmium (30.49 mg/kg of cadmium) reduced (P?<?0.05) the number of lymphocytes, the total erythrocyte count, the hemoglobin level, and the hematocrit. However, the activities of serum aspartate aminotransferase and gamma glutamyltransferase were increased (P?<?0.05). The total protein level was lower (P?<?0.05) in pigs fed with cadmium-contaminated diets. The contents of malondialdehyde increased (P?<?0.05), while the total antioxidant capacity and the activities of total superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and catalase decreased (P?<?0.05) in pigs fed with cadmium-contaminated diets. Dietary addition of maifanite can, to some extent, prevent the negative effects associated with feeding cadmium diets (30.49 mg/kg of cadmium) to pigs.     

Long-term in vitro culture of ovarian cortical tissue in goats: effects of FSH and IGF-I on preantral follicular development and FSH and IGF-I receptor mRNA expression

Long-term in vitro culture (16?days) of caprine ovarian cortical tissue was performed to test the effect of FSH and IGF-I on the viability and development of preantral follicles and mRNA expression for FSH and IGF-I receptors. Fragments were cultured in ??-MEM⁺ alone or supplemented with different combinations of FSH and IGF-I (sequential medium). The culture period was divided into two parts. Follicles were isolated and classified as normal or abnormal and primordial, primary or secondary. Viability of isolated follicles was determined by staining with Trypan Blue dye. Expression of FSHR and IGFR-1 mRNA was evaluated by qPCR. At day 8 of culture, more (P?<?0.05) follicles in treatments containing IGF-I alone or associated with FSH were normal and viable (overall mean, 81?% and 79?% respectively) than the treatments cultured with FSH or ??-MEM⁺ alone (68?% and 63?%). At day 16 of culture, treatments with FSH and/or IGF-I had more (P?<?0.05) viable follicles (69?%) than ??-MEM⁺ (38?%). The percentages of follicular development observed in the IGF-I/FSH, FSH+IGF-I/FSH+IGF-I and FSH/IGF-I treatments were similar but higher (P?<?0.05) than the other treatments. FSH and IGF-I during the entire culture period maximized (P?<?0.05) follicular and oocyte diameters and the percentage of secondary follicles (28?%). FSHR mRNA expression in the non-cultured control was similar to the treatment supplemented with FSH and IGF-I but higher (P?<?0.05) than ??-MEM⁺. IGFR-1 expression did not differ among treatments. Association of FSH and IGF-I in long-term in vitro culture promoted follicular development, maintaining FSHR mRNA expression.     

The Effectiveness of Cryoprotectants in Inhibiting Multiple Freeze-Thaw-Induced Functional and Rheological Changes in the Myofibrillar Proteins of Common Carp (Cyprinus carpio) Surimi

The purpose of the present study was to investigate the effects of cryoprotectants (4 % sucrose?+?4 % sorbitol) on functional and rheological changes, including thermal stability, dynamic rheological and emulsifying properties, gel texture profile, gel whiteness, gel water-binding capacity, and gel microstructure, in the myofibrillar proteins (MP) of common carp (Cyprinus carpio) surimi subjected to multiple freeze-thaw cycles (FT, 0, 1, 3 and 5 times). Increased numbers of FT cycles were accompanied by reduced thermal transition temperature (T _max), reduced enthalpy of denaturation (ΔH) (P?<?0.05), and enhanced susceptibility to thermal aggregation. The observation of the gel microstructure indicated that the FT processes produced empty spaces and changed the aggregate gel structure into a coarser-stranded network. The addition of cryoprotectants significantly prevented the reduction of MP thermal stability (P?<?0.05). The protective effect on protein functionality was demonstrated by the cryoprotectant’s efficacy in maintaining the three-dimensional gel network by small-strain oscillatory rheological testing and increasing the emulsifying capacity and emulsion stability of the MP. The FT process caused a decrease (P?<?0.05) in whiteness, water-binding capacity, and texture (hardness, springiness, and gumminess) of the MP gels, which were reduced by the addition of cryoprotectants. Overall, the FT-induced loss of the abovementioned functional and rheological properties was significantly reduced by the presence of cryoprotectants.     

Mitochondrial Complex Enzyme Activities and Cytochrome c Expression Changes in Multiple Sclerosis

Blood platelets have been widely proposed as biomarkers in studies of mitochondrial function and aging-related and neurodegenerative diseases. Defects in mitochondrial function were found not only in the substantia nigra of Parkinson’s disease patients but also in their blood platelets. Similarly, it has also been described in the blood platelet mitochondria of Alzheimer’s disease patients. To study mitochondrial aerobic metabolism function and protein expression in platelets of multiple sclerosis (MS) patients and control subjects, mitochondrial aconitase, mitochondrial superoxide dismutases 1 and 2 (SOD1 and SOD2), and respiratory complex enzyme activities in platelets of MS patients and control subjects were determined. Likewise, mitochondrial lipid peroxidation and mitochondrial SOD1 and cytochrome c expressions were investigated. Mitochondrial aconitase activity was higher in MS patients than in controls (P?<?0.05). A significant increase on all respiratory complex activities in MS patients was observed (P?<?0.05). Mitochondrial lipid peroxidation was significantly higher in MS patients than in controls (P?<?0.05). Significant changes of cytochrome c and mitochondrial SOD1 expressions were detected (P?<?0.05), with a decrease of 44?±?5 % and an increase of 46?±?6 %, respectively. Our study reveals that significant changes in mitochondrial aerobic metabolism function and mitochondrial SOD1 and cytochrome c expressions are produced in platelets of MS patients.     

Association of ADAM33 gene polymorphisms with asthma in Mongolian and Han groups in Inner Mongolia

Polymorphisms in the gene encoding for A disintegrin and metalloprotease 33 (ADAM33) are closely associated with the risk of bronchial asthma attacks in different populations. We collected blood samples from 248 asthma patients – 130 of the Han ethnic group and 118 of the Mongolian ethnic group – living in the Inner Mongolia region of China, and analyzed the single nuclear polymorphisms (SNPs) of the T1, T2 and V4 loci of the ADAM33 gene using PCR-RFLP (restriction fragment length polymorphism). In addition, we also tested 256 healthy controls (134 and 122 from the Han and Mongolian ethnic groups respectively) for the same SNPs. Three genotypes of the T1, T2 and V4 loci were predominantly detected: while polymorphisms in the T1 locus were significantly associated with asthma risk in both Mongolian and Han ethnicities (P?<?0.05, 1P?<?0.05), that in the V4 locus were relevant only in the Mongolian patients (P?<?0.05, 1P?>?0.05). In contrast, polymorphisms in the T2 locus showed no significant association with asthma risk in either ethnic group (P?>?0.05, 1P?>?0.05).     

Bayesian logistic regression in detection of gene–steroid interaction for cancer at <Emphasis Type="Italic">PDLIM5</Emphasis> locus

The PDZ and LIM domain 5 (PDLIM5) gene may play a role in cancer, bipolar disorder, major depression, alcohol dependence and schizophrenia; however, little is known about the interaction effect of steroid and PDLIM5 gene on cancer. This study examined 47 single-nucleotide polymorphisms (SNPs) within the PDLIM5 gene in the Marshfield sample with 716 cancer patients (any diagnosed cancer, excluding minor skin cancer) and 2848 noncancer controls. Multiple logistic regression model in PLINK software was used to examine the association of each SNP with cancer. Bayesian logistic regression in PROC GENMOD in SAS statistical software, ver. 9.4 was used to detect gene–steroid interactions influencing cancer. Single marker analysis using PLINK identified 12 SNPs associated with cancer (P < 0.05); especially, SNP rs6532496 revealed the strongest association with cancer (P = 6.84 × 10^?3); while the next best signal was rs951613 (P = 7.46 × 10^?3). Classic logistic regression in PROC GENMOD showed that both rs6532496 and rs951613 revealed strong gene–steroid interaction effects (OR = 2.18, 95% CI = 1.31?3.63 with P = 2.9 × 10^?3 for rs6532496 and OR = 2.07, 95% CI = 1.24 ?3.45 with P = 5.43 × 10^?3 for rs951613, respectively). Results from Bayesian logistic regression showed stronger interaction effects (OR = 2.26, 95% CI = 1.2 ?3.38 for rs6532496 and OR = 2.14, 95% CI = 1.14 ?3.2 for rs951613, respectively). All the 12 SNPs associated with cancer revealed significant gene–steroid interaction effects (P < 0.05); whereas 13 SNPs showed gene–steroid interaction effects without main effect on cancer. SNP rs4634230 revealed the strongest gene–steroid interaction effect (OR = 2.49, 95% CI = 1.5 ?4.13 with P = 4.0 × 10^?4 based on the classic logistic regression and OR = 2.59, 95% CI = 1.4 ?3.97 from Bayesian logistic regression; respectively). This study provides evidence of common genetic variants within the PDLIM5 gene and interactions between PLDIM5 gene polymorphisms and steroid use influencing cancer.     

Effects of Iron Glycine Chelate on Growth, Tissue Mineral Concentrations, Fecal Mineral Excretion, and Liver Antioxidant Enzyme Activities in Broilers

The study was conducted to determine the effects of iron glycine chelate (Fe-Gly) on growth, tissue mineral concentrations, fecal mineral excretion, and liver antioxidant enzyme activities in broilers. A total of 360 1-day-old commercial broilers (Ross?×?Ross) were randomly allotted to six dietary treatments with six replications of ten chicks per replicate. Broilers were fed a control diet with no Fe supplementation, while five other treatments consisted of 40, 80, 120, and 160?mg Fe/kg diets from Fe-Gly, and 160?mg Fe/kg from ferrous sulfate, respectively. After a 42-day feeding trial, the results showed that 120 and 160?mg Fe/kg as Fe-Gly improved the average daily gain (P?<?0.05) and average daily feed intake (P?<?0.05) of broilers (4?C6?weeks). Addition with 120 and 160?mg Fe/kg from Fe-Gly and 160?mg Fe/kg from FeSO₄ increased Fe concentration in serum (P?<?0.05), liver (P?<?0.05), breast muscle (P?<?0.05), tibia (P?<?0.05), and feces (P?<?0.01) at 21 and 42?days. There were linear responses to the addition of Fe-Gly from 0 to 160?mg/kg Fe on Fe concentration in serum (21?days, P?=?0.005; 42?days, P?=?0.001), liver (P?=?0.001), breast muscle (P?=?0.001), tibia (P?=?0.001), and feces (21?days, P?=?0.011; 42?days, P?=?0.032). Liver Cu/Zn superoxide dismutase activities of chicks were increased by the addition of 80, 120, and 160?mg Fe/kg as Fe-Gly to diets at 42?days. There were no differences in liver catalase activities of chicks among the treatments (P?>?0.05). This study indicates that addition with Fe-Gly could improve growth performance and iron tissue storage and improves the antioxidant status of broiler chickens.     

Effects of dietary l-arginine or N-carbamylglutamate supplementation during late gestation of sows on the miR-15b/16, miR-221/222, VEGFA and eNOS expression in umbilical vein

Placental vascular formation and blood flow are crucial for fetal survival, growth and development, and arginine regulates vascular development and function. This study determined the effects of dietary arginine or N-carbamylglutamate (NCG) supplementation during late gestation of sows on the microRNAs, vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) expression in umbilical vein. Twenty-seven landrace?×?large white sows at day (d) 90 of gestation were assigned randomly to three groups and fed the following diets: a control diet and the control diet supplemented with 1.0% l-arginine or 0.10% NCG. Umbilical vein of fetuses with body weight around 2.0?kg (oversized), 1.5?kg (normal) and 0.6?kg (intrauterine growth restriction, IUGR) were obtained immediately after farrowing for miR-15b, miR-16, miR-221, miR-222, VEGFA and eNOS real-time PCR analysis. Compared with the control diets, dietary Arg or NCG supplementation enhanced the reproductive performance of sows, significantly increased (P?<?0.05) plasma arginine and decreased plasma VEGF and eNOS (P?<?0.05). The miR-15b expression in the umbilical vein was higher (P?<?0.05) in the NCG-supplemented group than in the control group. There was a trend in that the miR-222 expression in the umbilical vein of the oversized fetuses was higher (0.05?<?P?<?0.1) than in the normal and IUGR fetuses. The expression of eNOS in both Arg-supplemented and NCG-supplemented group were lower (P?<?0.05) than in the control group. The expression of VEGFA was higher (P?<?0.05) in the NCG-supplemented group than in the Arg-supplemented and the control group. Meanwhile, the expression of VEGFA of the oversized fetuses was higher (P?<?0.05) than the normal and IUGR fetuses. In conclusion, this study demonstrated that dietary Arg or NCG supplementation may affect microRNAs (miR-15b, miR-222) targeting VEGFA and eNOS gene expressions in umbilical vein, so as to regulate the function and volume of the umbilical vein, provide more nutrients and oxygen from the maternal to the fetus tissue for fetal development and survival, and enhance the reproductive performance of sows.     

Effects of Selenium Source and Level on Growth Performance, Tissue Selenium Concentrations, Antioxidation, and Immune Functions of Heat-Stressed Broilers

An experiment is conducted to investigate the effects of selenium (Se) source and level on growth performance, tissue Se concentrations, antioxidation, and immune functions of heat-stressed broilers from 22 to 42?days of age. A total of 210 22-day-old Arbor Acres commercial male chicks were assigned by body weight to one of seven treatments with six replicates of five birds each in a completely randomized design involving a 3?×?2 factorial arrangement plus one Se-unsupplemented basal diet control (containing 0.027?mg of Se/kg). The three Se sources were sodium selenite (Na₂SeO₃), Se yeast, and AMMS Se (Se protein), and the two supplemental Se levels were 0.15 or 0.30?mg Se/kg. All birds were reared under heat-stressed condition (33?±?1?°C during 0900?C1700?hours and 27?±?1?°C during 1900?C0700?hours with a relative humidity of 60?C80?%). The results showed that heat-stressed chicks fed Se-supplemented diets had higher (P?<?0.10) average daily feed intake, Se concentrations in liver and breast muscle, liver glutathione peroxidase (GSH-Px) activity, serum antibody titers against H5N1(Re-4 strain), H5N1(Re-5 strain) and lower (P?<?0.01) mortality compared with the control. Chicks fed the diets supplemented with 0.30?mg/kg of Se had higher (P?<?0.05) Se concentrations in liver and breast muscle, liver GSH-Px activity, and serum antibody titer against H5N1 (Re-4 strain) than those fed the diets supplemented with 0.15?mg/kg of Se. Broilers fed the diets supplemented with Se yeast had higher (P?<?0.001) Se concentrations in liver and breast muscle than those fed the diets supplemented with Na₂SeO₃ or AMMS Se. However, broilers fed the diets supplemented with AMMS Se had higher (P?<?0.05) serum antibody titers against H5N1 (Re-4 strain) and H5N1 (Re-5 strain) than those fed the diets supplemented with Na₂SeO₃. These results indicated that Se yeast was more effective than Na₂SeO₃ or AMMS Se in increasing tissue Se retention; however, AMMS Se was more effective than Na₂SeO₃ or Se yeast in improving immune functions of heat-stressed broilers.