bESC from cloned embryos do not retain transcriptomic or epigenetic memory from somatic donor cells.

In brief: Epigenetic reprogramming after mammalian somatic cell nuclear transfer is often incomplete, resulting in low efficiency of cloning. However, gene expression and histone modification analysis indicated high similarities in transcriptome and epigenomes of bovine embryonic stem cells from in vitro fertilized and somatic cell nuclear transfer embryos. Abstract: Embryonic stem cells (ESC) indefinitely maintain the pluripotent state of the blastocyst epiblast. Stem cells are invaluable for studying development and lineage commitment, and in livestock, they constitute a useful tool for genomic improvement and in vitro breeding programs. Although these cells have been recently derived from bovine blastocysts, a detailed characterization of their molecular state is lacking. Here, we apply cutting-edge technologies to analyze the transcriptomic and epigenomic landscape of bovine ESC (bESC) obtained from in vitro fertilized (IVF) and somatic cell nuclear transfer (SCNT) embryos. bESC were efficiently derived from SCNT and IVF embryos and expressed pluripotency markers while retaining genome stability. Transcriptome analysis revealed that only 46 genes were differentially expressed between IVF- and SCNT-derived bESC, which did not reflect significant deviation in cellular function. Interrogating histone 3 lysine 4 trimethylation, histone 3 lysine 9 trimethylation, and histone 3 lysine 27 trimethylation with cleavage under targets and tagmentation, we found that the epigenomes of both bESC groups were virtually indistinguishable. Minor epigenetic differences were randomly distributed throughout the genome and were not associated with differentially expressed or developmentally important genes. Finally, the categorization of genomic regions according to their combined histone mark signal demonstrated that all bESC shared the same epigenomic signatures, especially at gene promoters. Overall, we conclude that bESC derived from SCNT and IVF embryos are transcriptomically and epigenetically analogous, allowing for the production of an unlimited source of pluripotent cells from high genetic merit organisms without resorting to transgene-based techniques.

Systematic alteration of ATAC-seq for profiling open chromatin in cryopreserved nuclei preparations from livestock tissues.

The use of Assay for Transposase-Accessible Chromatin (ATAC-seq) to profile chromatin accessibility has surged over the past years, but its applicability to tissues has been very limited. With the intent of preserving nuclear architecture during long-term storage, cryopreserved nuclei preparations from chicken lung were used to optimize ATAC-seq. Sequencing data were compared with existing DNase-seq, ChIP-seq, and RNA-seq data to evaluate library quality, ultimately resulting in a modified ATAC-seq method capable of generating high quality chromatin accessibility data from cryopreserved nuclei preparations. Using this method, nucleosome-free regions (NFR) identified in chicken lung overlapped half of DNase-I hypersensitive sites, coincided with active histone modifications, and specifically marked actively expressed genes. Notably, sequencing only the subnucleosomal fraction dramatically improved signal, while separation of subnucleosomal reads post-sequencing did not improve signal or peak calling. The broader applicability of this modified ATAC-seq technique was tested using cryopreserved nuclei preparations from pig tissues, resulting in NFR that were highly consistent among biological replicates. Furthermore, tissue-specific NFR were enriched for binding motifs of transcription factors related to tissue-specific functions, and marked genes functionally enriched for tissue-specific processes. Overall, these results provide insights into the optimization of ATAC-seq and a platform for profiling open chromatin in animal tissues.

A sexual intimacy survey of former nuns and priests.

Men and women who have lived in a celibate religious community experience a unique set of sexual, social, and psychological problems upon resuming a secular life style. In many instances the personality factors and circumstances which led both to a decision to enter and then to leave a celibate religious community are not easily appreciated by the nonreligious professional counselor and do not readily lend themselves to extrapolation from other population groups. This article reports the findings of a preliminary study to identify the sexual experiences and problems of persons who have left religious communities. Data is based on responses to a mailed, anonymous questionnaire by 126 former nuns and priests living in all parts of the country. Information is reported on the following areas: (1) sexual behavior and enjoyment prior to, while living in, and after leaving a religious community; (2) current sexual behavior, satisfaction and problems; (3) sexual counseling experience; and (4) general problems and concerns with integrating sexual intimacy into present life styles. Comparisons are made with similar data published in the literature and data obtained from persons attending Sexual Attitude Reassessment (SAR) Workshops. The findings of this study suggest that persons who have left celibate religious communities have had limited sexual experience in adolescence and early adulthood compared to nonreligious persons. Further, there are special sexual problems and needs in readjusting to a secular life style. Implications for specific counseling are discussed.

Human sexuality: an interdisciplinary program for health care professionals and the physically disabled.

The central feature of a program in human sexuality for health care professionals and persons with physical disabilities developed at the Texas Institute for Rehabilitation and Research (TIRR) is a 2 1/2-day Sexual Attitude Reassessment (SAR) Workshop. Although the content of this Workshop has many features in common with ones used elsewhere, its organization, financing, and development are relatively unique. Based in a small, specialty hospital in the South, it is a low-budget, largely self-supporting program run by an extradepartmental, multidisciplinary staff. Evaluations of 12 workshops involving over 500 disabled and ablebodied participants are compared with similar data reported in the literature along with information (before and after the workshop) concerning the participants' attitudes, behavior, and satisfaction regarding various sexual activities. There were programmatic, institutional, and community problems unique to developing this program, but it has unusual potential for personal and staff development, intrainstitutional team building and broader community service.