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 method for visualizing CellML models.

MOTIVATION: The Physiome Project was established in 1997 to develop tools to facilitate international collaboration in the physiological sciences and the sharing of biological models and experimental data. The CellML language was developed to represent and exchange mathematical models of biological processes. CellML models can be very complicated, making it difficult to interpret the underlying physical and biological concepts and relationships captured/described in the mathematical model. RESULTS: To address this issue a set of ontologies was developed to explicitly annotate the biophysical concepts represented in the CellML models. This article presents a framework that combines a visual language, together with CellML ontologies, to support the visualization of the underlying physical and biological concepts described by the mathematical model and also their relationships with the CellML model. Automated CellML model visualization assists in the interpretation of model concepts and facilitates model communication and exchange between different communities.

Biophysical annotation and representation of CellML models.

MOTIVATION: CellML is an implementation-independent model description language for specifying and exchanging biological processes. The focus of CellML is the representation of mathematical formulations of biological processes. The language captures the mathematical and model building constructs well, but does not lend itself to capturing the biology these models represent. RESULTS: This article describes the development of an ontological framework for annotating CellML models with biophysical concepts. We demonstrate that, by using these ontological mappings, in combination with a set of graph reduction rules, it is possible to represent the underlying biological process described in a CellML model.

Facilitating modularity and reuse: guidelines for structuring CellML 1.1 models by isolating common biophysical concepts.

The CellML language was developed in response to the need for a high-level language to represent and exchange mathematical models of biological processes. The flexible structure of CellML allows modellers to construct mathematical models of the same biological system in many different ways. However, some modelling styles do not naturally lead to clear abstractions of the biophysical concepts and produce CellML models that are hard to understand and from which it is difficult to isolate parts that may be useful for constructing other models. In this article, we advocate building CellML models which isolate common biophysical concepts and, using these, to build mathematical models of biological processes that provide a close correspondence between the CellML model and the underlying biological process. Subsequently, models of higher complexity can be constructed by reusing these modularized CellML models in part or in whole. Development of CellML models that best describe the underlying biophysical concepts thus avoids the need to code models from scratch and enhances the extensibility, reusability, consistency and interpretation of the models.

The evolution of CellML.

CellML is an open XML-based markup language for describing and exchanging mathematical models of biological processes. While CellML was originally designed to describe and exchange models of cellular and subcellular processes, the design principles that were applied to the construction of a language with this relatively narrow focus are equally applicable to the specification of a language with a much wider scope. In this paper we describe the structure of CellML, how the language is evolving to broaden its scope, and what tools are being developed to facilitate its use.

Predictors of coping efficacy, negative moods and post-traumatic stress syndrome following major trauma.

The purpose of the study was to test relationships among injury appraisals, coping, social support and outcomes of coping efficacy, mood states and post-traumatic stress syndrome. A purposive sample of 152 hospitalized patients a week following major trauma was recruited from two trauma centres. The measures included the modified Ways of Coping Scale; the brief Social Support Questionnaire; the brief Profile of Mood State; the Impact of Event Scale; and selected demographic and injury related variables. Data were analyzed by multivariate statistics. The results indicated that perceived controllability, satisfaction with social support, wishful-thinking coping and problem-focused coping were the significant predictors for coping efficacy; perceived stressfulness, problem-focused coping and wishful thinking explained a significant amount of variance in mood states; avoidance coping and wishful-thinking coping were predictors for post-traumatic stress syndrome. The moderating effects of emotion-focused coping at high and medium levels were identified.

Struggling with paradoxes: the process of spiritual development in women with cancer.

PURPOSE/OBJECTIVES: To examine the process of spiritual development in women diagnosed with cancer within five years of initial treatment. DESIGN: Exploratory, qualitative. SETTING: Outpatients in the mid-central and southwestern United States. SAMPLE: 10 Caucasian women, ages 45-70, who completed initial treatment, were not undergoing treatment for recurrence, and were within five years of diagnosis for breast or ovarian cancer or non-Hodgkin's lymphoma. METHODS: Data collected during two semistructured interviews, coded and analyzed using grounded theory techniques. Frame of reference--symbolic interactionism. MAIN RESEARCH VARIABLES: Developmental processes of spirituality; responses to diagnosis, treatment, and survival of cancer. FINDINGS: Diagnosis of cancer threatened the meaning of the women's lives, resulting in a sense of disintegration. This problem was resolved through the basic social psychological process of Struggling With Paradoxes, a three-phase process of Deciphering the Meaning of Cancer for Me, Recognizing Human Limitations, and Learning to Live with Uncertainty. In phase I, the paradoxes focused on the possibility of death, distress, vulnerability, and maintaining connection. In phase II, the paradoxes involved confronting death, asking difficult questions, and letting go of ultimate control of their lives. In phase III, the paradoxes centered on uncertainty, redefining meaning, and identifying spiritual growth. Reintegration occurred over time, although when threatened by the possibility of recurrence, disintegration resurfaced for a time. CONCLUSIONS: Findings emphasize not only the importance of spirituality, but also that spiritual experience is individualized and developmental in nature. Spiritual growth occurs over time following the diagnosis of cancer and is not necessarily related to age. IMPLICATIONS FOR NURSING PRACTICE: Spiritual concerns may be painful for patients to address; spiritual caregiving requires an acknowledgment of need by the woman with cancer and a caring, sensitive caregiver. Nurses should be aware of the phases of spiritual development so that interventions can be designed to address individual needs that may vary over time.

Transformation of peripheral inputs by the first-order lateral line brainstem nucleus.

Extracellular recording techniques were used to record the responses of medial nucleus cells and posterior lateral line nerve fibers in mottled sculpin, Cottus bairdi, and goldfish, Carassius auratus, to a 50-Hz dipole source (vibrating sphere). Responses were characterized in terms of (1) receptive fields that relate responsiveness (spike rate and phase-locking) to the location of the source along the length of the fish, (2) input-output functions that relate responsiveness to vibration amplitude for a fixed source location, and (3) peri-stimulus time histograms that relate responsiveness to time during a sustained period of vibration. Relative to posterior lateral line nerve fibers, medial nucleus cells in both species were similar in showing (1) lower spontaneous and evoked rates of spike activity, (2) greater degrees of adaptation, (3) greater heterogeneity in all response characteristics, and (4) evidence for inhibitory/excitatory interactions. Whereas receptive fields of nerve fibers in both species faithfully reflect both pressure gradient amplitudes (with rate changes) and directions (with phase-angle changes) in the stimulus field, receptive fields of medial nucleus were more difficult to relate to the stimulus field, Some, but not all, receptive fields could be modeled with excitatory center/inhibitory surround and inhibitory center/excitatory surround organizations.

Attempting to fathom the unfathomable: descriptive views of spirituality.

OBJECTIVES: To examine the various views of "spirituality and its uses in the disciplines of philosophy, theology, psychology, and nursing. DATA SOURCES: Definitions and descriptions of spirituality and related terms from the disciplines of philosophy, theology, psychology, and nursing. CONCLUSION: Although it is widely accepted that holistic nursing care incorporates care of the spirit, nursing's view of spirituality is influenced by varying paradigms. Nursing researchers are exploring spirituality. Spiritual care that is ethical and sensitive is an invaluable part of total patient care. IMPLICATIONS FOR NURSING PRACTICE: Appreciating various views of spirituality and recognizing the possible discrepancy between a nurse's and a patient's view of spirituality allows the reader to use such terms carefully and appropriately in providing sensitive patient care.

Hindbrain signal processing in the lateral line system of the dwarf scorpionfish Scopeana papillosus

Recordings were made from primary afferent fibres and secondary projection neurones (crest cells) in the mechanosensory lateral line system of the dwarf scorpionfish. Crest cells were identified by antidromic stimulation from the contralateral midbrain. Differences between primary afferent fibre and crest cell response characteristics are indicative of signal processing by the neuronal circuitry of the medial octavolateralis nucleus. There are a number of differences between primary afferent fibres and crest cells. Primary afferents have relatively high levels of spontaneous activity (mean close to 40 impulses s-1) and many of them are strongly modulated by ventilation. By contrast, crest cells have a much lower rate of spontaneous activity that is not obviously modulated by ventilation. Primary afferents show a simple tonic response to a maintained stimulus, whereas crest cells show a variety of temporal response properties, but in general show a phasic/tonic response to the same prolonged stimulus. Afferents are most sensitive to frequencies of stimulation around 100 Hz; in contrast, crest cells show a strong suppression of activity at this frequency. Crest cells are most responsive around 50 Hz. These afferent/secondary comparisons show similarities with those reported for allied electrosensory and auditory pathways.

A spline surface algorithm for reconstruction of corneal topography from a videokeratographic reflection pattern.

A videokeratograph quantifies one or more aspects of corneal shape by the computer analysis of a digital image. The image is the reflection formed by illuminating the cornea with a known source pattern. The type of shape information recovered and the accuracy of this information depend upon properties of the computer algorithm used in the analysis. We present a new algorithm that recovers a polynomial spline description of corneal depth.

Coping strategies of long-term cancer survivors.

Cancer survival is a stressful experience requiring coping for the maintenance of equilibrium. Lazarus' Theory of Stress and Coping was the framework for this descriptive study of the use and effectiveness of coping strategies as assessed by long-term survivors of cancer. The Jalowiec Coping Scale (JCS) and a subject information sheet (SIS) were mailed to 128 potential subjects, identified by the snowball technique, who survived cancer for > 5 years, were not currently receiving therapy, and were not in a terminal stage of disease. Fifty-nine subjects with a mean survival of 13.03 years correctly completed and returned the questionnaire and were included in data analysis. Respondents were predominantly white (88.1%), female (83.7%), married (72.8%), employed as professionals (57.8%), 41-65 years of age (59.3%), and diagnosed with breast cancer (50.8%). Subjects rated optimistic, supportive, and confrontive strategies as most often used and effective. Length of survival did not result in different choices of strategies. Statistically significant differences were found in coping styles between elderly and middle-aged survivors. Results of this study increase nurses' awareness of effective coping strategies and the importance of assessment of coping in long-term survivors of cancer. The importance of social support, spirituality, and helping others is emphasized.

Glucose transporters and in vivo glucose uptake in skeletal and cardiac muscle: fasting, insulin stimulation and immunoisolation studies of GLUT1 and GLUT4.

Our aim was to study glucose transporters GLUT1 and GLUT4 in relation to in vivo glucose uptake in rat cardiac and skeletal muscle. The levels of both transporters were of a similar order of magnitude in whole muscle tissue (GLUT1/GLUT4 ratio varied from 0.1 to 0.6), suggesting that both may have an important physiological role in regulating muscle glucose metabolism. GLUT4 correlated very strongly (r2 = 0.97) with maximal insulin-stimulated glucose uptake (Rg' max., estimated using the glucose clamp plus 2-deoxy[3H]glucose bolus technique) in six skeletal muscles and heart. A distinct difference in regulation of the two transporters was evident in heart: in 5 h-fasted rats, basal glucose uptake and GLUT1 levels in heart were very high and both were reduced, by 90 and 60% respectively, by 48 h fasting. However, in heart (and in red skeletal muscle), neither GLUT4 levels nor Rg' max. were reduced by 48 h fasting. GLUT1 was shown to be specifically expressed in cardiac myocytes, because intracellular vesicles enriched in GLUT4 contained significant levels of GLUT1. In conclusion, the high association of muscle GLUT4 content with insulin responsiveness in different muscles, and the preservation of both with fasting, supports a predominant role of GLUT4 in insulin-mediated glucose uptake. GLUT1 may play an important role in mediating cardiac muscle glucose uptake in the basal metabolic state. Marked changes in GLUT1 expression with alterations in the metabolic state, such as prolonged fasting, may play an important role in cardiac glucose metabolism.

Prostanoids and hypothermic renal preservation injury.

The effect of 48 hours of hypothermic renal ischemia utilizing Euro-Collins flush and short term reperfusion on renal prostaglandin synthesis was studied in dogs. Hypothermic ischemia followed by 60 minutes of reperfusion in-vivo resulted in significant elevations in renal Thromboxane B2 (TXB2) production in the outer cortex, inner cortex, and medulla, relative to non-ischemic kidneys. Prostaglandin E2 (PGE2) and 6-keto Prostaglandin F1 alpha (6-K PGF1 alpha) production were not significantly affected by ischemia and reperfusion. Enhanced TXB2 production was not seen with ischemia alone (without reperfusion) or with reperfusion with O2 saturated buffer, indicating a blood born source or stimuli. Early postreperfusion renal blood flow after hypothermic ischemia followed a biphasic pattern; blood flow increased for the first 10 minutes of reperfusion to achieve normal values, and then steadily declined over the next 20 minutes. This pattern was not altered by the cyclooxygenase inhibitors Idomethacin (5 mg/kg, P.O.) or Mefenamic acid (10 mg/kg, I.V.). Administration of the TXA2 synthesis inhibitor CGS-12970 (3 mg/kg, I.V.) or the TXA2/endoperoxide receptor antagonist SQ-29548 (80 micrograms/min, I.A.) significantly increased renal blood flow during reperfusion but neither agent altered the basic time dependent pattern observed in the control group. These data indicate that 48 hours of hypothermic renal ischemia results in dramatic changes in intrarenal TXA2 synthesis at the time of reperfusion. Enhanced TXA2 production is not dependent on reoxygenation per se, but rather requires reperfusion with blood suggesting a circulatory source.(ABSTRACT TRUNCATED AT 250 WORDS)

Personal safety. It is a matter of awareness.

The safety and security points discussed in this article are only a few of the awareness tips that can add to your personal safety. When reporting to work, especially during odd hours, be suspicious of all activities. Ask yourself, "What if?" Becoming a victim or not becoming a victim is easy.

Sexual attitudes, behavior and satisfaction for able-bodied and disabled participants attending workshops in human sexuality.

Since 1973, Sexual Attitude Reassessment (SAR) Workshops have been conducted for persons with physical handicaps, health care professionals and interested members of the community. Although reassessment of personal sexual attitudes is the primary objective of the workshops, several closely related objectives have emerged: (1) evaluation of the impact of the experience on changing both sexual attitudes and behaviors; (2) evaluation of the sexual satisfaction and problem areas of participants with particular reference to persons with physical disabilities; and (3) provision of an institute-wide catalyst for initiating programs and services based on newly identified unmet needs. This article presents data from 15 workshops with emphasis on comparing the sexual attitudes, behavior and satisfaction of the able-bodied and the disabled participants. Of the 650 participants, 98 (15%) were disabled and of this group approximately 75% had spinal cord injury. Comparison of pre- and post-workshop responses to sexual attitude scales showed similar and significant changes on four of nine scales for both the able-bodied and disabled groups. Analysis of sexual behavior and satisfaction by physical status revealed that the disabled participants were less active than the able-bodied participants although both groups reported similar percentages of satisfaction (38 and 41% respectively) with their current level of sexual activity. Lack of partners was cited most frequently by both groups as the major reason for dissatisfaction with current sexual status. Implications of these findings are discussed with regard to sexual counseling and areas of further research.

A family practice residency inpatient service: a review of 631 admissions.

This study reports a review of 631 admissions by family practice residents and staff over a 12-month period to an autonomous family practice service in a large US Army medical center. The diagnoses, number and types of consultations requested, types of patients cared for by residents in various levels of taining, and other pertinent data are reviewed. The study indicates that an inpatient family practice service can be very successful in terms of physician and patient satisfaction.