Genome-wide association study and genomic heritabilities for blood protein levels in Lori-Bakhtiari sheep.

Serum protein levels are related to physiological and pathological status of animals and could be affected by both genetic and environmental factors. This study aimed to evaluate genetic variation of serum protein profile in sheep. Blood samples were randomly collected from 96 Lori-Bakhtiari ewes, a heavy meat-type breed. Total protein, albumin, globulin, α1, α2, ß and γ globulins and IgG levels were measured in blood serum. The samples were genotyped using the Illumina OvineSNP50 BeadChip. The studied traits adjusted for age, birth type, birth season and estimate of breeding value for body weight were considered as pseudo-phenotypes in genome-wide association analysis. In the GWAS model, the first five principal components were fitted as covariates to correct the biases due to possible population stratification. The Plink, R and GCTA software were used for genome-wide association analysis, construction of Q-Q and Manhattan plots and estimation of genetic variances, respectively. Noticeable genomic heritabilities ± SE were estimated for total and γ globulins (0.868 ± 0.262 and 0.831 ± 0.364, respectively), but other protein fractions had zero or close to zero estimates. Based on the Bonferroni adjusted p values, four QTLs located on 181.7 Mbp of OAR3, 107.7 Mbp of OAR4, 86.3 Mbp of OAR7 and 83.0 Mbp of OAR8 were significantly associated with α1, ß, ß and γ globulins, respectively. The results showed that the PKP2, IGF2R, SLC22A1 and SLC22A2 genes could be considered as candidate genes for blood serum proteins. The present study showed significant genetic variations of some blood protein fractions.

Genome-wide association study of four composite reproductive traits in Iranian fat-tailed sheep.

Composite reproductive traits are a combination of growth and reproductive traits. They have the advantage of being better attuned to the market drivers since producers are paid on a per kilogram basis and not on a per head basis. In this study, 124 Lori---Bakhtiari ewes were genotyped using the medium-density Illumina Ovine SNP50 array. A genome-wide association study was performed on estimated breeding values of four composite reproductive traits and genetic parameters were also estimated. The traits were litter mean weight at birth, litter mean weight at weaning, total litter weight at birth and total litter weight at weaning. Several suggestive and associated single nucleotide polymorphisms (SNPs) were identified. Neighbouring the top SNPs there were five genes, inhibin ß E subunit (INHBE), inhibin ß C subunit (INHBC), testis expressed 12 (TEX12), ß-carotene oxygenase 2 (BCO2) and WD Repeat Domain 70 (WDR70) identified as possible candidate genes for composite reproductive traits of the Lori-Bakhtiari sheep. These genes are in pathways known to be relevant to fertility and growth characteristics. The results provide new information for the functional annotation of genes associated with fertility traits and add new evidence towards a consensus of quantitative trait loci associated with reproductive traits in sheep.

Genome-wide association study of birth weight in sheep.

Birth weight is the earliest available growth trait with considerable impacts on lamb survivability and growth performance traits. This study was conducted to perform a genome-wide association study of birth weight in a meat-type sheep. A total of 132 Lori-Bakhtiari sheep were selected based on estimated of breeding values (EBVs) for BW analyses. The selected animals were genotyped using Illumina Ovine SNP50 Bead Chip. After quality control, a total of 41 323 single-nucleotide polymorphisms (SNPs) and 130 sheep were used for subsequent analyses. Plink 1.90 beta software was used for the analyses. Seven SNPs on chromosomes 1, 16, 19 and 22 were detected based on genome-wide unadjusted P-values (P <10-6), which jointly accounted for 1.2% of total genetic variation. However, based on Bonferroni-adjusted P-values, only three SNPs on chromosome 1 had significant associations with EBVs for birth weight (P <0.05), which jointly explained 0.8% of total genetic variation. A total of seven genes were found in 50 kb intervals from the three significant SNPs on chromosome 1, but only three genes, including RAB6B (a member of RAS oncogene family), Tf serotransferrin and GIGYF2 (a GRB10 interacting GYF protein 2), could be considered as candidate genes for birth weight in future studies. The results of this study may facilitate potential use of the genes involving in growth and production traits for genetic improvement of productivity in sheep.

Genome-wide association study to identify genomic regions affecting prolificacy in Lori-Bakhtiari sheep.

Several causative mutations in candidate genes affecting prolificacy have been detected in various sheep breeds. A genome-wide association study was performed on estimated breeding values for litter size in Lori-Bakhtiari sheep. Prolific ewes with twinning records and others with only singleton records were genotyped using the medium-density Illumina Ovine SNP50 array. Four single nucleotide polymorphisms (SNPs) associated with litter size were identified on chromosomes 3, 6 and 22. The region on sheep chromosome 3 between 75 739 167 and 75 745 152 bp included two significant SNPs (s52383.1 and OAR3_80038014_X.1) in high linkage disequilibrium with each other. The region that surrounds these SNPs contains a novel putative candidate gene: luteinizing hormone/choriogonadotropin receptor (LHCGR), known to be involved in ovarian steroidogenesis and organism-specific biosystem pathways in sheep. Known prolificacy genes BMPR1B, BMP15 and GDF9 were not associated with litter size in Lori-Bakhtiari sheep, suggesting that other biological mechanisms could be responsible for the trait's variation in this breed.

Assessment of narrative writing by Persian-speaking students with hearing impairments.

OBJECTIVE: Previous studies have highlighted that narrative skill is critical to the development of the literacy skills by children. Children with cochlear implants (CI) and hearing aids (HA) may have problems in narrative development compared to peers with healthy hearing (HH). There is no exact data about the narrative writing ability of Persian-speaking students who are hearing-impaired. This study was undertaken to compare the microstructure and macrostructure scores for narrative writing of Persian-speaking students who are hearing-impaired and peers with HH. DESIGN: This was a cross-sectional descriptive-analytical study. SETTING: The subjects were recruited from elementary schools in the city of Tehran. PARTICIPANTS: A total of 144 elementary school students were participated. OUTCOME MEASURES: The written narratives were elicited using a wordless pictorial storybook story. Three-way ANOVA with post hoc adjusted Bonferroni test was applied to determine the main effects and interactions of grounded variables on the microstructure and macrostructure components of narrative writing. RESULTS: No significant differences were observed in the macrostructure components of narrative writing between hearing-impaired and HH students. Factors analysis showed that the 4th grade HH students had significantly the highest scores, and the 3rd grade HA students had significantly the lowest scores in microstructure components of narrative writing. CONCLUSION: The findings revealed that hearing-impaired students similarly to their HH peers can transmit the main idea (macrostructure) of narrative writing, but show critical difficulties when using complete grammatical elements (microstructures) to form sentences to convey the idea in the narrative.

Correlation of Gankyrin oncoprotein overexpression with histopathological grade in prostate cancer.

Prostate cancer is the second most commonly diagnosed cancer among men worldwide. Identifying new prognostic and predictive biomarkers will help stratification of prostate cancer patients for a better treatment. Gankyrin is a novel oncoprotein which regulates cell cycle and protein degradation. Gankyrin overexpression correlated with the malignant phenotypes and promotes the tumorigenicity and metastasis in many cancers. However, there are not any reports on the role of Gankyrin in prostate cancer. Therefore, this study was designed to investigate the expression of Gankyrin in prostate cancer and analyze its correlation with some clinicopathological characteristics. We characterized the expression of gankyrin in fifty five prostate cancer specimens and twenty non-cancerous tissues by immunohistochemical staining and the results were correlated with clinical characteristics and pathological parameters.Results showed that Gankyrin was expressed in 41 of 55 (74%) prostate cancer patients and its expression was significantly higher than corresponding adjacent normal tissues (p<0.001). Gankyrin overexpression was significantly correlated with histopathological tumor grade, Gleason score and tumor differentiation (P=0.002). These findings showed that Gankyrin is mainly overexpressed in high grade prostate tumors so it may have a significant role in prostate cancer progress and it May serve as a useful biomarker for the identification of aggressive prostate cancers.

Dominance genetic and maternal effects for genetic evaluation of egg production traits in dual-purpose chickens.

1. A study was conducted to study direct dominance genetic and maternal effects on genetic evaluation of production traits in dual-purpose chickens. The data set consisted of records of body weight and egg production of 49 749 Mazandaran fowls from 19 consecutive generations. Based on combinations of different random effects, including direct additive and dominance genetic and maternal additive genetic and environmental effects, 8 different models were compared. 2. Inclusion of a maternal genetic effect in the models noticeably improved goodness of fit for all traits. Direct dominance genetic effect did not have noticeable effects on goodness of fit but simultaneous inclusion of both direct dominance and maternal additive genetic effects improved fitting criteria and accuracies of genetic parameter estimates for hatching body weight and egg production traits. 3. Estimates of heritability (h2) for body weights at hatch, 8 weeks and 12 weeks of age (BW0, BW8 and BW12, respectively), age at sexual maturity (ASM), average egg weights at 28-32 weeks of laying period (AEW), egg number (EN) and egg production intensity (EI) were 0.08, 0.21, 0.22, 0.22, 0.21, 0.09 and 0.10, respectively. For BW0, BW8, BW12, ASM, AEW, EN and EI, proportion of dominance genetic to total phenotypic variance (d2) were 0.06, 0.08, 0.01, 0.06, 0.06, 0.08 and 0.07 and maternal heritability estimates (m2) were 0.05, 0.04, 0.03, 0.13, 0.21, 0.07 and 0.03, respectively. Negligible coefficients of maternal environmental effect (c2) from 0.01 to 0.08 were estimated for all traits, other than BW0, which had an estimate of 0.30. 4. Breeding values (BVs) estimated for body weights at early ages (BW0 and BW8) were considerably affected by components of the models, but almost similar BVs were estimated by different models for higher age body weight (BW12) and egg production traits (ASM, AEW, EN and EI). Generally, it could be concluded that inclusion of maternal effects (both genetic and environmental) and, to a lesser extent, direct dominance genetic effect would improve the accuracy of genetic evaluation for early age body weights in dual-purpose chickens.

A review on prolificacy genes in sheep.

Ovulation rate and litter size are important reproduction traits in sheep and are of high economic value. Reproduction traits typically have low to medium heritabilities and do not exhibit a noticeable response to phenotypic selection. Therefore, inclusion of genetic information of the genes associated with reproductive ability could efficiently enhance the selection response. The most important major genes affecting prolificacy and their genetic diversities in different sheep breeds were reviewed. Different causative mutations with major effects on reproductive traits including ovulation rate and litter size have been found in various sheep breeds around the world. A general overview of the studies on main prolificacy genes showed that some alleles may express different phenotypic effects in different breeds, and thus, further studies on epistatic effects are necessary for more understanding of genetic control of reproductivity in sheep. Regarding the polygenic control of fertility traits, application of new high-throughput technologies to find new variants is essential for future studies. Moreover, genomewide association studies and genomic best linear unbiased predictions of breeding values are likely to be effective tools for genetic improvement of sheep reproductive performance traits.

Novel Mutation in Exon 1 of the BMP15 Gene and its Association with Reproduction Traits in Sheep.

The BMP15 gene is a growth factor and a member of the transforming growth factor ß (TGFß) superfamily, specifically expressed in oocytes. In the present study, polymorphism of BMP15 gene exon 1 was studied using single strand conformational polymorphism (SSCP) and direct DNA sequencing methods in 170 Mehraban and Lori sheep ewes. A 231-bp fragment in BMP15 exon 1 was amplified by PCR reactions. Two genotypes (GG and AG) with a new point mutation at position 121 bp of the studied fragment (c.379G>A in reference GenBank number AF236078.1 sequence), deducing an amino acid exchange in the codified amino acid sequence (p.Glu41Lys) were identified in the studied populations. The AG and GG frequencies were 74.4% and 25.6% in Mehraban and 44.7% and 55.3% in Lori sheep, respectively. Frequencies of the A and G alleles were 37.2% and 62.8% in Mehraban and 22.4% and 77.6% in Lori sheep, respectively. Two different secondary structures of protein were predicted for encoded precursor protein. The genotypes GG and AG did not have any significant association with the studied reproductive traits, but the AA genotype is likely to have a lethal or sterility effect.

Comparison of different models for genetic evaluation of egg weight in Mazandaran fowl.

1. The aim of the present study was to compare different models to estimate variance components for egg weight (EW) in laying hens. 2. The data set included 67 542 EW records of 18 245 Mazandaran hens at 24, 28, 30, 32 and 84 weeks of age, during 19 consecutive generations. Variance components were estimated using multi-trait, repeatability, fixed regression and random regression models (MTM, RM, FRM and RRM, respectively) by Average Information-Restricted Maximum Likelihood algorithm (AI-REML). The models were compared based on Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC). 3. The MTM was the best model followed by the Legendre RRMs. A RRM with 2nd degree of fit for fixed regression and 3(rd) and 2(nd) degrees of fit for random regressions of direct additive genetic and permanent environmental effects, respectively, was the best RRM. The FRM and RM were not proper models to fit the data. However, nesting curves within contemporary groups improved the fit of FRM. 4. Heritability estimates for EW by MTM (0.06-0.41) were close to the estimates obtained by the best RRM (0.09-0.45). In both MTM and RRM, positive genetic correlations were estimated for EW records at different ages, with higher correlations for adjacent records. 5. The results suggest that MTM is the best model for EW data, at least when the records are taken at relatively few age points. Though selection based on EW at higher ages might be more precise, 30 or 32 weeks of age could be considered as the most appropriate time points for selection on EW to maximise genetic improvement per time unit.

SNP exploring in the middle and terminal regions of the IGF-1 gene and association with production and reproduction traits in Holstein cattle.

Five primer sets were designed in order to identify single nucleotide polymorphisms (SNPs) in middle and terminal exons (2 to 6) and in some flanking intronic regions of the bovine insulin-like growth factor 1 (IGF-1) gene. Sequencing results of PCR products for 10% of animals showed no variant in exons but a SNP at intron 4 was occurred. Both polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and high resolution melting (HRM) methods were developed to genotype samples. The PCR-RFLP results showed the presence of three fragments on agarose gel for the C allele due to two cleavage sites while two fragments for the T allele were observed. Melting curves of 123bp fragments in HRM analysis showed a difference between temperature melting (Tm) of two homozygous genotypes as the CC genotypes had higher Tm than the TT genotypes. Melting curve of the CT genotype was different and crossed two parallel patterns of homozygous genotypes. The frequencies of the CC, CT and TT genotypes were 0.6, 0.37 and 0.03, respectively. Also, the estimated allele frequencies were 0.785 and 0.215 for the C and T alleles, respectively. Results showed higher accuracy of the HRM analysis compared to the PCR-RFLP method. Least square means (LSMs) comparison of the different genotypes in the SNP showed significant association with milk fat yield trait in the first lactation and open days after the second calving. The polymorphism did not have a significant effect on other milk production or reproduction traits. It seems that other variants or QTLs known in this region underlie genetic variation in the production and reproduction of dairy cattle.

Association of BMPR-1B and GDF9 genes polymorphisms and secondary protein structure changes with reproduction traits in Mehraban ewes.

BMPR-1B and GDF9 genes are well known due to their important effects on litter size and mechanisms controlling ovulation rate in sheep. In the present study, polymorphisms of BMPR-1B gene exon 8 and GDF9 gene exon 1 were detected by single strand conformational polymorphism (SSCP) analysis and DNA sequencing methods in 100 Mehraban ewes. The PCR reaction forced to amplify 140 and 380-bp fragments of BMPR-1B and GDF9 genes, respectively. Two single nucleotide polymorphisms (SNPS) were identified in two different SSCP patterns of BMPR-1B gene (CC and CA genotypes) that deduced one amino acid exchange. Also, two SNPS were identified in three different SSCP patterns of GDF9 gene (AA, AG and GG genotypes) that deduced one amino acid exchanges. Two different secondary structures of protein were predicted for BMPR-1B exon 8, but the secondary protein structures predicted for GDF9 exon 1 were similar together. The evaluation of the associations between the SSCP patterns and the protein structure changes with reproduction traits showed that BMPR-1B exon 8 genotypes have significant effects on some of reproduction traits but the GDF9 genotypes did not have any significant effect. The CA genotype of BMPR-1B exon 8 had a significant positive effect on reproduction performance and could be considered as an important and new mutation, affecting the ewes reproduction performance. Marker assisted selection using BMPR-IB gene could be noticed to improve the reproduction traits in Mehraban sheep.

Effect of Dietary Cation-Anion Difference during Prepartum and Postpartum Periods on Performance, Blood and Urine Minerals Status of Holstein Dairy Cow.

Twenty four periparturient cows were used to determine the effects of DCAD on acid-base balance, plasma and urine mineral concentrations, health status, and subsequent lactation performance. Each group of 12 cows received either a diet containing -100 DCAD or +100 DCAD for 21 d prepartum. Both anionic and cationic groups were divided into two groups, one received a +200 DCAD and the other +400 DCAD diet for 60 d postpartum. Prepartum reduction of DCAD decreased DMI, urinary and blood pH, urinary concentrations of Na or K and increased plasma and urinary Ca, Mg, Cl and S. Also cows fed -100 DCAD diet consumed the most dry matter in the first 60 d after calving. Postpartum +400 DCAD increased milk fat and total solid percentages, urinary and blood pH and urinary Na and K concentrations, but urinary Ca, P, Cl and S contents decreased. Greater DMI, FCM yields were observed in cows fed a diet of +400 DCAD than +200 DCAD. No case of milk fever occurred for any diets but feeding with a negative DCAD diet reduced placenta expulsion time. In conclusion, feeding negative DCAD in late gestation period and high DCAD in early lactation improves performance and productivity of dairy cows.

Comparison of different models for estimation of genetic parameters of early growth traits in the Mehraban sheep.

Genetic parameters for birth weight (BW), weaning weight (WW) and pre-weaning daily gain (PWDG) in Iranian Mehraban sheep were estimated using restricted maximum likelihood (REML) procedure. Six different animal models were fitted, differentiated by including or excluding maternal effects, with and without covariance between maternal and direct genetic effects. The estimates for direct heritability ranged from 0.26 to 0.53, 0.18 to 0.32 and 0.15 to 0.33 for BW, WW and PWDG respectively. The estimates were substantially higher when maternal effects, either genetic or environmental, were ignored in the model. The results of this study show that full models with maternal genetic and environmental effects gave the most accurate estimates for early growth traits.

High-dose methylprednisolone for children with acute lymphoblastic leukemia and unfavorable presenting features.

In an attempt to improve treatment outcome high-dose methylprednisolone (HDMP, 20-30 mg/kg, once a day orally) was used instead of a conventional dose of steroid (2 mg/kg/d, in 3 divided doses) in children with acute lymphoblastic leukemia (ALL) with increased risk factors. HDMP combined with cytotoxic agents (vincristine and L-asparaginase) resulted in an improved complete remission rate (94%) in 48 newly diagnosed children with ALL compared to 81% in 86 historical controls receiving standard dose steroid combined with the same treatment regimen. The bone marrow relapse rate was lower in patients who received HDMP (31%) than in controls (56%). Treatment was discontinued in 56% of 48 patients receiving HDMP and in 35% of 86 controls. The difference was significant (p < 0.05). The 5-yr continuous complete remission rate was significantly greater in patients received HDMP compared with the control patients (60% vs. 43%, p < 0.05). HDMP treatment was well tolerated without significant adverse effects. Moreover, during induction therapy the duration of leukopenia (< 2 x 10(9)/L) was shorter in patients receiving HDMP. We conclude that HDMP combined with other antileukemic agents increased the CR rate and prolonged the duration of remission in children with ALL who had increased risk factors. However, the optimal dosage of HDMP and its role in maintenance therapy should be determined in future, randomized studies.

[Effect of surgical draping on bacterial contamination in the surgical field]. / Einfluss der Inzisionsfolie auf das Keimverhalten im Operationsgebiet.

Standardized samples were taken from the skin of 100 patients immediately before and after their hip operation. 50 patients were operated with and 50 patients without use of plastic drapes. The samples were evaluated quantitatively and qualitatively by the employment of usual microbiological methods. The statistical analysis revealed that at the end of the operations there were a) as many microbes on the drape as on the skin where no drape had been used and b) many more microbes under the drapes than either on them or on undraped skin. The value of the plastic drape as a means of preventing infections has to be doubted. Discontinuing the use of plastic drapes seems to be justified.