Nearly 7% of men are afflicted by male infertility worldwide, and genetic factors are suspected to play a significant role in the majority of these patients. Although sperm morphology is an important parameter measured in the semen analysis, only a few genetic causes of teratozoospermia are currently known. The objective of this study was to define the association between alterations in the genes encoding the Golgi-associated PDZ- and coiled-coil motif containing protein (GOPC), the protein interacting with C kinase 1 (PICK1) and the acrosomal protein zona pellucida binding protein 1 (ZPBP1/sp38) with abnormal sperm head morphology in infertile men. Previous reports demonstrated that mice lacking Gopc, Pick1 and Zpbp1 are infertile due to abnormal head morphology. Herein, using our validated RNA-based method, we studied spermatozoal cDNA encoding the human GOPC, PICK1 and ZPBP1 genes in 381 teratozoospermic and 240 controls patients via direct sequencing. Among these genes, we identified missense and splicing mutations in the sperm cDNA encoding ZPBP1 in 3.9% (15/381) of men with abnormal sperm head morphology. These mutations were not observed in 240 matched controls and the dbSNP database (χ(2) = 9.3, P = 0.002). In contrast, statistically significant and functionally relevant mutations were not discovered in the GOPC and PICK1 genes. In our study ZPBP1 mutations are associated with abnormal sperm head morphology, defined according to strict criteria, resembling the mouse Zpbp1 null phenotype. We hypothesize that missense mutations exert a dominant-negative effect due to altered ZPBP1 protein folding and protein:protein interactions in the acrosome.

Background The H19/IGF2 imprinted loci have attracted recent attention due to their role in cellular differentiation and proliferation, heritable gene regulation, and in utero or early postnatal growth and development. Expression from the imprinted H19/IGF2 locus involves a complex interplay of three means of epigenetic regulation: proper establishment of DNA methylation, promoter occupancy of CTCF and expression of microRNA-675 (miR675). We have previously demonstrated in a multigenerational rat model of intrauterine growth restriction the epigenetic heritability of adult metabolic syndrome in a F2 generation. We have further demonstrated abrogation of the F2 adult metabolic syndrome phenotype with essential nutrient supplementation of intermediates along the one-carbon pathway, and shown that alterations in the metabolome precede the adult onset of metabolic syndrome. However, the upstream molecular and epigenomic mediators underlying these observations have yet to be fully elucidated. Objective In the current study, we sought to characterize the impact of the intrauterine growth restricted lineage and essential nutrient supplementation on both levels and molecular mediators of H19 and IGF2 gene expression in the F2 generation. Study Design F2 intrauterine growth restricted and sham lineages were obtained by exposing P1 (grandmaternal) pregnant dams to bilateral uterine artery ligation or sham surgery at gestational day 19.5. F1 pups were allocated to the essential nutrient supplemented or control diet at postnatal day 21, and bred at 6–7 weeks of age. Hepatic tissues from the resultant F2 offspring at birth and at weaning (day 21) were obtained. Bisulfite modification and sequencing was employed for methylation analysis. H19 and IGF2 expression was measured by QPCR. Promoter occupancy was quantified using chromatin immunoprecipitation, or ChIP, against CTCF insulator proteins. Results Growth-restricted F2 on control diet demonstrated significant down-regulation in H19 expression as compared to sham lineage (0.7831 vs 1.287; p< 0.05); however, essential nutrient supplementation diet abrogates this difference (4.995 vs 5.100; p>0.05). Conversely, Igf2 was up regulated by essential nutrient supplemented diet on the sham lineage (2.0 fold, p=0.01), an effect that was not observed in the growth restricted offspring. A significant differential methylation was observed in the promoter region of region H19 among the intrauterine growth restricted lineage (18% vs 25%; p<0.05) on a control diet, while the essential nutrient supplemented diet was alternately associated with hypermethylation in both lineages (sham: 50%; IUGR: 84%, p<0.05). Consistent with essential nutrient supplementation impacting the epigenome, a decrease of CTCF promoter occupancy was observed in CTCF4 of the growth restricted lineage (2.45% vs 0.56%; p<0.05) on the control diet, an effect that was repressed with essential nutrient supplementaion. Conclusions Heritable growth restriction is associated with changes in H19 gene expression; the...

SummaryIn our primate model of maternal high fat diet exposure, we have described fetal epigenomic modifications to the peripheral circadian Npas2 are associated with persistent alterations in fetal hepatic metabolism and non-alcoholic fatty liver. As the interaction of circadian response with metabolism is not well understood, we employed a murine knockout model to characterize the molecular mechanisms with which Npas2 reprograms the fetal hepatic metabolic response. cDNA was generated from Npas2 -/-and +/+ (wild type) livers at day 2 (newborn) and at 25 weeks (adult) of life. Newborn samples were analyzed by exon array (n=3/cohort). Independent pathway analysis software determined that the primary dysregulated pathway(s) in the Npas2 -/-animals uniformly converged on lipid metabolism. Of particular interest, Ppargc1a, which integrates circadian and metabolism pathways, was significantly (p<.01) over expressed in newborn (1.7 fold) and adult (1.8 fold) Npas2-/-animals. These findings are consistent with an essential role for Npas2 in programming the peripheral circadian response and hepatic metabolism, which has not been previously described.

Background We have recently shown in both non-human primates and in rodents that fetal and neonatal hepatic expression of the circadian transcription factor, Npas2, is modulated by a high fat maternal diet and plays a critical role in establishing life-long metabolic homeostasis. Similarly, we and others have also established the importance of the maternal and early postnatal diet on establishment of the early gut microbiome. Objective We hypothesized that altered circadian gene expression solely in the neonatal liver would result in gut microbiome dysbiosis, especially with diet-induced metabolic stress (i.e., restricted feeding). Using a murine model where we conditionally knock out of Npas2 in the neonatal liver, we aimed to determine the role of the circadian machinery in gut dysbiosis with restricted feeding. Design We collected fecal samples from liver Npas2 conditional knock out (cKO; n=11) and wild type (wt: n=13) reproductive-aged mice before (study day 0) and after the restricted feeding study (study day 17). Extracted DNA was sequenced using the MiSeq Illumina platform using primers specific for the V4 region of the 16S rDNA gene. The resulting sequences were quality filtered, aligned, and assigned taxonomy. Principal coordinate analysis was performed on unweighted and weighted UniFrac distances between samples with Permutation ANOVA to assess clustering significance between groups. Microbial taxa that significantly differ between groups of interest was determined using Linear Discriminate Analysis Effect Size and randomForrest. Results Principal coordinate analysis performed on weighted UniFrac distances between male cKO and wt cohorts revealed that the gut microbiome of the mice did not differ by genotype at the start of the restricted feeding study, but did differ by virtue of genotype at the end of the study (p=0.001). Moreover, these differences could be at least partially attributed to restricted feeding-associated alterations in relative abundance of the Bacteroides genus, which has been implicated as crucial to establishing a healthy gut microbiome early in development. Conclusions Here we have provided an initial key insight into the interplay between neonatal establishment of the peripheral circadian clock in the liver and the ability of the gut microbiome to respond to dietary and metabolic stress. As Npas2 expression in the liver is a target of maternal high fat diet induced metabolic perturbations during fetal development, we speculate that these findings have potential implications in the long term metabolic health of their offspring.

This study proposes a strategy development framework for information management in a construction business. The framework covers the typical project lifecycle of a construction business which includes work winning and delivery. It incorporates a cultural analysis approach that covers the range of cultural manifestations expected in organisations. The approach also combines qualitative and quantitative research techniques. Application of the proposed framework for the development of information management strategy in a leading construction business showed that examining the information culture of the leadership and staff of the organisation is helpful in exposing the relevant informational issues. The case study also helped to validate that the proposed framework is a viable method for identifying and resolving informational issues inherent in construction organisations. Lastly, considering that the information management strategy developed in this study is for a typical construction business, other companies in the same domain will find the resource valuable.