Mammalian Glutaminase Gls2 Gene Encodes Two Functional Alternative Transcripts by a Surrogate Promoter Usage Mechanism

Glutaminase is expressed in most mammalian tissues and cancer cells, but the regulation of its expression is poorly understood. An essential step to accomplish this goal is the characterization of its species- and cell-specific isoenzyme pattern of expression. Our aim was to identify and characterize transcript variants of the mammalian glutaminase Gls2 gene.We demonstrate for the first time simultaneous expression of two transcript variants from the Gls2 gene in human, rat and mouse. A combination of RT-PCR, primer-extension analysis, bioinformatics, real-time PCR, in vitro transcription and translation and immunoblot analysis was applied to investigate GLS2 transcripts in mammalian tissues. Short (LGA) and long (GAB) transcript forms were isolated in brain and liver tissue of human, rat and mouse. The short LGA transcript arises by a combination of two mechanisms of transcriptional modulation: alternative transcription initiation and alternative promoter. The LGA variant contains both the transcription start site (TSS) and the alternative promoter in the first intron of the Gls2 gene. The full human LGA transcript has two in-frame ATGs in the first exon, which are missing in orthologous rat and mouse transcripts. In vitro transcription and translation of human LGA yielded two polypeptides of the predicted size, but only the canonical full-length protein displayed catalytic activity. Relative abundance of GAB and LGA transcripts showed marked variations depending on species and tissues analyzed.This is the first report demonstrating expression of alternative transcripts of the mammalian Gls2 gene. Transcriptional mechanisms giving rise to GLS2 variants and isolation of novel GLS2 transcripts in human, rat and mouse are presented. Results were also confirmed at the protein level, where catalytic activity was demonstrated for the human LGA protein. Relative abundance of GAB and LGA transcripts was species- and tissue-specific providing evidence of a differential regulation of GLS2 transcripts in mammals

Ensuring competency in end-of-life care: controlling symptoms

BACKGROUND: Palliative medicine is assuming an increasingly important role in patient care. The Education for Physicians in End-of-life Care (EPEC) Project is an ambitious program to increase core palliative care skills for all physicians. It is not intended to transmit specialty level competencies in palliative care. METHOD: The EPEC Curriculum was developed to be a comprehensive syllabus including trainer notes, multiple approaches to teaching the material, slides, and videos of clinical encounters to trigger discussion are provided. The content was developed through a combination of expert opinion, participant feedback and selected literature review. Content development was guided by the goal of teaching core competencies not included in the training of generalist and non-palliative medicine specialist physicians. RESULTS: Whole patient assessment forms the basis for good symptom control. Approaches to the medical management of pain, depression, anxiety, breathlessness (dyspnea), nausea/vomiting, constipation, fatigue/weakness and the symptoms common during the last hours of life are described. CONCLUSION: While some physicians will have specialist palliative care services upon which to call, most in the world will need to provide the initial approaches to symptom control at the end-of-life

Perda da espontaneidade da ação: o desconforto de homens que sofreram infarto agudo do miocardio Perdida de la expontaneidad de la accion: el desconforto de hombres que sufrieron infarto agudo del miocardio Undergoing loss of spontaneous action: the discomfort experienced by men having suffered acute myocardial infarction

Considerando que conforto - cuja promoção é uma das metas centrais da enfermagem -- e desconforto devem ser entendidos àluz das interações vivenciadas pelo paciente - e, portanto, vinculadas aos fatores objetivos das instituições, da nacionalidade epráticas que as fundamentam - este estudo investigou os significados de conforto e desconforto na perspectiva de homens quesofreram infarto agudo do miocárdio (IAM). Recorrendo ao Interacionismo Simbólico eàmetodologia da Teoria Fundamentadanos Dados, entrevistas foram feitas com 13 homens que sofreram IAM, em duas unidades de saúde na cidade de São Paulo. Aanálise permitiu construir um modelo teórico daqueles significados nessa experiência, caracterizada por três fenômenos eessencialmente permeada pelo desconforto de "padecer a perda da espontaneidade daação ".Este artigo resume o processopsicossocial básico da experiência, discutindo suas implicações para questionar o modelo clínico de intervenção, apontando apossibilidade de a enfermagem atuar na prevenção e propondo questões para aprimorar a formação do enfermeiro.<br>Considerando que conforto - cuya promoción es una de las metas centralés de la enfermeria - y desconforto deven ser entendidos a la luz de las interacciones vivenciadas por el paciente e por lo tanto, vinculadas a los factores objetivos de las instituciones, dela racionalidad y de las prácticas que las fundamentan - este estudio investigo los significados de conforto e desconforto en la prespectiva de hombres que sufrieron infarto agudo del miocardio (IAM). Acudiendo al Interaccionismo SimbólicoYala meto-dologia de la Teoria Fudamentada en los datos, fueron realizadas entrevistas com 13 hombres que sufrieron IAM en dos unida-des de salud en la cuidad de São Paulo. El análisis permitio construir um modelo teórico de aquellos significados en esa experi-encia, caracterizada por tres fenómenos y esencialmente condicionada por el desconforto de "padecer la pérdida de la exponta-neidad de la accion". Este artículo, resume el proceso psicosocial básico de la experiencia, discutiendo sus implicancias paracuestionar el modelo clínico de intervención, apuntando la posibilidad dela enfermera para actuar en la prevención y proponerideas para aprimorar la formación del enfermero.<br>Considering that comfort and discomfort must be understood in the light of patients' interactions during illness and treatment- thus linked to institutions' objective factors, grounding rationale, and practices -, this study inquired on comfort and discom-fort such as experienced by men who had suffered acute myocardial infarction (AMI). By resorting to the Symbolic Interactionismand to Grounded Theory methodology, data were collected by means of interviews with 13 men who had suffered AMI, at twohealth units in the city of São Paulo. The analysis led to building a theoretical model of such experience, made up by threephenomena and essentially pervaded by the discomfort of "undergoing loss of spontaneous action". This articles sums up thebasic psychosocial process that emerges from that experience, and discusses its implications to question the clinical model oftreatment, pointing to prevention as a further scope of action for nurses, besides raising issues to enhance nurse education