Transcription factor Zbtb1 interacts with bridging factor Lmo2 and maintains the T-lineage differentiation capacity of lymphoid progenitor cells.

Hematopoietic stem and progenitor cells can differentiate into all types of blood cells. Regulatory mechanisms underlying pluripotency in progenitors, such as the ability of lymphoid progenitor cells to differentiate into T-lineage, remain unclear. We have previously reported that LIM domain only 2 (Lmo2), a bridging factor in large transcriptional complexes, is essential to retain the ability of lymphoid progenitors to differentiate into T-lineage. However, biochemical characterization of Lmo2 protein complexes in physiological hematopoietic progenitors remains obscure. Here, we identified approximately 600 Lmo2-interacting molecules in a lymphoid progenitor cell line by two-step affinity purification with LC-MS/MS analysis. Zinc finger and BTB domain containing 1 (Zbtb1) and CBFA2/RUNX1 partner transcriptional corepressor 3 (Cbfa2t3) were found to be the functionally important binding partners of Lmo2. We determined CRISPR/Cas9-mediated acute disruption of Zbtb1 or Cbfa2t3 in the lymphoid progenitor or bone marrow-derived primary hematopoietic progenitor cells causes significant defects in the initiation of T-cell development when Notch signaling is activated. Our transcriptome analysis of Zbtb1- or Cbfa2t3-deficient lymphoid progenitors revealed that Tcf7 was a common target for both factors. Additionally, ChIP-seq analysis showed that Lmo2, Zbtb1, and Cbfa2t3 cobind to the Tcf7 upstream enhancer region, which is occupied by the Notch intracellular domain/RBPJ transcriptional complex after Notch stimulation, in lymphoid progenitors. Moreover, transduction with Tcf7 restored the defect in the T-lineage potential of Zbtb1-deficient lymphoid progenitors. Thus, in lymphoid progenitors, the Lmo2/Zbtb1/Cbfa2t3 complex directly binds to the Tcf7 locus and maintains responsiveness to the Notch-mediated inductive signaling to facilitate T-lineage differentiation.

Transcription factors regulate early T cell development via redeployment of other factors: Functional dynamics of constitutively required factors in cell fate decisions.

Establishment of cell lineage identity from multipotent progenitors is controlled by cooperative actions of lineage-specific and stably expressed transcription factors, combined with input from environmental signals. Lineage-specific master transcription factors activate and repress gene expression by recruiting consistently expressed transcription factors and chromatin modifiers to their target loci. Recent technical advances in genome-wide and multi-omics analysis have shed light on unexpected mechanisms that underlie more complicated actions of transcription factors in cell fate decisions. In this review, we discuss functional dynamics of stably expressed and continuously required factors, Notch and Runx family members, throughout developmental stages of early T cell development in the thymus. Pre- and post-commitment stage-specific transcription factors induce dynamic redeployment of Notch and Runx binding genomic regions. Thus, together with stage-specific transcription factors, shared transcription factors across distinct developmental stages regulate acquisition of T lineage identity.

Notch signaling supports the appearance of follicular helper T cells in the Peyer's patches concomitantly with the reduction of regulatory T cells.

The intracellular fragment of Notch1, a mediator of Notch signaling that is frequently detected in thymic immigrants, is critical for specifying T-cell fate in the thymus, where Delta-like 4 (Dll4) functions as a Notch ligand on the epithelium. However, as such Notch signaling has not been detected in mature T cells, how Notch signaling contributes to their response in secondary lymphoid organs has not yet been fully defined. Here, we detected the marked expression of Dll4 on the stromal cells and the active fragment of Notch1 (Notch1 intracellular domain, N1ICD) in CD4+ T cells in the follicles of Peyer's patches (PPs). In addition, N1ICD-bearing T cells were found in the T-cell zone of PPs, especially in the transcription factor Foxp3+ regulatory T (Treg) cells, with slight expression of Dll4 on the stromal cells. These fragments disappeared in Dll4-deficient conditions. It was also found that Notch1- and Notch2-deficient T cells preferentially differentiated into Treg cells in PPs, but not CXCR5+PD-1+ follicular helper T (Tfh) cells. Moreover, these phenotypes were also observed in chimeric mice reconstituted with the control and T-cell-specific Notch1/2-deficient bone marrow or Treg cells. These results demonstrated that Dll4-mediated Notch signaling in PPs is required for the efficient appearance of Tfh cells in a Treg cell-prone environment, which is common among the gut-associated lymphoid tissues, and is critical for the generation of Tfh-mediated germinal center B cells.

LMO2 is essential to maintain the ability of progenitors to differentiate into T-cell lineage in mice.

Notch signaling primarily determines T-cell fate. However, the molecular mechanisms underlying the maintenance of T-lineage potential in pre-thymic progenitors remain unclear. Here, we established two murine Ebf1-deficient pro-B cell lines, with and without T-lineage potential. The latter expressed lower levels of Lmo2; their potential was restored via ectopic expression of Lmo2. Conversely, the CRISPR/Cas9-mediated deletion of Lmo2 resulted in the loss of the T-lineage potential. Introduction of Bcl2 rescued massive cell death of Notch-stimulated pro-B cells without efficient LMO2-driven Bcl11a expression but was not sufficient to retain their T-lineage potential. Pro-B cells without T-lineage potential failed to activate Tcf7 due to DNA methylation; Tcf7 transduction restored this capacity. Moreover, direct binding of LMO2 to the Bcl11a and Tcf7 loci was observed. Altogether, our results highlight LMO2 as a crucial player in the survival and maintenance of T-lineage potential in T-cell progenitors via the regulation of the expression of Bcl11a and Tcf7.

Stage-specific action of Runx1 and GATA3 controls silencing of PU.1 expression in mouse pro-T cells.

PU.1 (encoded by Spi1), an ETS-family transcription factor with many hematopoietic roles, is highly expressed in the earliest intrathymic T cell progenitors but must be down-regulated during T lineage commitment. The transcription factors Runx1 and GATA3 have been implicated in this Spi1 repression, but the basis of the timing was unknown. We show that increasing Runx1 and/or GATA3 down-regulates Spi1 expression in pro-T cells, while deletion of these factors after Spi1 down-regulation reactivates its expression. Leveraging the stage specificities of repression and transcription factor binding revealed an unconventional but functional site in Spi1 intron 2. Acute Cas9-mediated deletion or disruption of the Runx and GATA motifs in this element reactivates silenced Spi1 expression in a pro-T cell line, substantially more than disruption of other candidate elements, and counteracts the repression of Spi1 in primary pro-T cells during commitment. Thus, Runx1 and GATA3 work stage specifically through an intronic silencing element in mouse Spi1 to control strength and maintenance of Spi1 repression during T lineage commitment.

Notch2 complements Notch1 to mediate inductive signaling that initiates early T cell development.

Notch signaling is the dominant intercellular signaling input during the earliest stages of T cell development in the thymus. Although Notch1 is known to be indispensable, we show that it does not mediate all Notch signaling in precommitment stages: Notch2 initially works in parallel to promote early murine T cell development and antagonize other fates. Notch-regulated target genes before and after T lineage commitment change dynamically, and we show that this partially reflects shifts in genome-wide DNA binding by RBPJ, the transcription factor activated by complex formation with the Notch intracellular domain. Although Notch signaling and transcription factor PU.1 can activate some common targets in precommitment T progenitors, Notch signaling and PU.1 activity have functionally antagonistic effects on multiple targets, delineating separation of pro-T cells from alternative PU.1-dependent fates. These results define a distinct mechanism of Notch signal response that distinguishes the initial stages of murine T cell development.

[Teaching-learning strategies in nursing--analysis using the Glasgow Coma Scale]. / Estratégias de ensino-aprendizagem na enfermagem: análise pela Escala de Coma de Glasgow.

Using the Glasgow Coma Scale (ECGI) as a subject, this paper aims to analyze and verify the apprehension of knowledge towards the teaching-learning and self-learning offered to nursing workers, and to check the degree of knowledge acquired during the process and possible results stemming from the nursing student/nursing worker association. This descriptive, quantitative-based study counted on the participation of 62 currently enrolled students in the first semester of the 4th year of nursing. The following teaching-learning strategies were used: expositive classes with the use of slides and videotape, and a basic text. Among participants, 41.9% were nursing workers; 61.3% informed to have taken care of patients with high alteration of the consciousness level, predominantly located in their working group. Statistically, there has been a successful improvement in the percentage of correct actions after the expositive class and videotape. Self-learning results showed no alteration either. Intergroup correlations displayed no disparity in the degree of acquired knowledge.

[Morbidity and mortality from external causes in Brazil, 2000]. / As causas externas no Brasil no ano 2000: comparando a mortalidade e a morbidade.

Since the early 1980s, external causes have been the second greatest cause of death in Brazil. They also place a growing demand on health care services. The current study analyzes morbidity and mortality from external causes in Brazil. The study material consists of 118,367 deaths and 652,249 hospitalizations due to external causes during the year 2000. The data are from the National Mortality Information System and Hospital Information System. The mortality coefficient from external causes was 69.7/ 100 thousand (119.0/100 thousand for males and 21.8/100 thousand for women). Homicides were the leading cause of death (38.3% of the total), with a high coefficient of 26.7/100 thousand, while falls were the leading cause of hospitalizations (42.8% of the total). Motor vehicle accidents were a major cause of both morbidity and mortality. Fractures, mostly occurring in the upper and lower limbs, accounted for 42.6% of hospitalizations. Based on the findings, preventive programs should aim to decrease both mortality and morbidity, with special emphasis on homicides, traffic accidents, and falls.

[Injury among the elderly: the challenge to integrate preventive activities in public and individual levels]. / Mortes e internações por causas externas entre os idosos no Brasil: o desafio de integrar a saúde coletiva e atenção individual.

BACKGROUND: In Brazil and all over the world the elderly are increasing as a result of the raise in life expectancy. This group still faces a significant risk for some diseases as well as injuries. The proposal of this study was to describe fatal and nonfatal injuries among people aged 60 years and older in Brazil. SETTING: Brazil, the last year available was 2000. METHODS: The data from 13,383 injury deaths and 87,177 outcomes among people aged 60 years and older were analyzed. The data sets were obtained from Federal Health Department of Brazil from Mortality System Information (from death certificates) and Hospitalization Information System (from discharges registered in public hospitals). RESULTS: The mortality rate is 92.1/100,000 (135.3/100,000 for male and 56.8/100,000 for female) that are higher than overall population rates, especially for women. The lead cause is transport accidents (27.5% of total injury deaths) which mortality rate is 25.3/100,000 and 48.2% are pedestrian. Homicides rate is 9.5/100,000, it is almost three times lower than for total population in Brazil. Falls rate is 14.0/100,000, it is the third place in injury deaths for men and women and suicide rate is 6.9/100,000, lower than developed countries. Opposite to mortality, falls are the major cause of nonfatal injuries hospitalization for both men and women, accounting for 48,940 discharges (56.1%). Fractures are 52.8% of all injuries, especially in falls and transport accidents. CONCLUSIONS: It is crucial to develop injury preventive activities in both public and individual level. Falls should receive emphasis in injury prevention efforts.

[Evaluation of conscious level in head-injury patients]. / AVALIAÇÃO NO NÍVEL DE CONSCIÊNCIA EM PACIENTES COM TRAUMATISMO CRÂNIO - ENCEFÁLICO.

A study regarding the evaluation of conscious level in 50 patients with head-injury, admitted in Neurosurgical Intensive Care Unit. For evaluation, the patient was observed and examined in his own bed, bee utilized a scale with three behaviors tables, a minimum score de 3 pointing the smaller degree of conscious level, and maximum score of 15 pointing the neurophysiologic normal state. The utilized method has demonstrated to be usefull for the precision facilities verified in evaluated answers and for the reduced time of appliance, which was around five minutes as a rule.

[Intracranial pressure monitoring]. / MONITORIZAÇÂO DA PRESSÃO INTRACRANIANA.

Basic aspects of intracranial pressure monitoring are presented. They are necessary for to effective action of the nurse who looks after patients submitted to this procedure.

[The nurse facing increased intracranial pressure patient]. / A ENFERMEIRA EM FACE DO PACIENTE COM HIPERTENSÃO INTRACRANIANA.

A review of increased intracranial pressure. Physiopathological aspects and preventive measures, which can be adopted by nurses, in the imminence of sudden increased intracranial pressure.

[Specialization course in intensive care nursing - part. I: student's prolife]. / CURSO DE ESPECIALIZAÇÃO DE CUIDADOS INTENSIVOS DE ENFERMAGEM ­ PARTE I ­ PERFIL DOS ALUNOS.

This is Part I of a two part papers. After completing five specialization courses in intensive care nursing, the authors report their experience in this Part and they describe the type of students all graduate nurses, who enrolled in such courses.

[Specialization course in intensive care nursing of "Escola de Enfermagem da USP" - part 2: the learning-teaching process]. / CURSO DE ESPECIALIZAÇÃO EM ENFERMAGEM DE CUIDADOS INTENSIVOS DA ESCOLA DE ENFERMAGEM DA USP ­ PARTE 2: O PROCESSO DE ENSINO-APRANDIZAGEM ADOTADO NO CURSO.

In this article, the authors describe the course's structure, methods and strategies applied to the learning-teaching process. Emphasis is given to the critical analyses and nursing research, within a proper environment for the exchange of experiences between teachers and students, looking for the improvement of professionals activing in intensive care units.

[Perception of Intensive Care Unit patients - problems felt and expectations in relation of nursing care]. / PERCEPÇÃO DOS PACIENTES DE UNIDADE DE TERAPIA INTENSIVA Problemas sentidos e expectativas em relação à assistência de enfermagem.

A survey made with adult patients, submitted to cardiac surgery, who stayed in Intensive Care Unit (ICU) between 24 to 72 hours. The main point of the survey was to verify the most frequent problems felt by the patients in ICU and their expectations in relation to nursin care. It was found that, the majority of the problems felt and of the expectations related to nursing assistance belong in the basic area, more so than in the technical area.

[Retrospective analysis of nursing research in critical care nursing: 1975 to 1984]. / ANÁLISE RETROSPECTIVA DAS PESQUISAS DE ENFERMAGEM EM TERAPIA INTENSIVA: 1975-1984.

Nursing research in Intensive Care Units as compered to nursing research in other medical-surgical units, both in Brasil and in the United States, is the subject of this study. Data were collected from articles published in 1975, 1979 and 1984 in "Revista Brasileira de Enfermagem" (for Brasil) and "Nursing Research" and "Heart and Lungs" (for USA). The analysis of data included type of study, its depth, metodology employed and selected subject. Increase in number of studies in nursing in Intensive Care Units, both descriptive and experimental, was observed. The authors also noted the absence of replication of research and of continuity studies.

[Loss of spontaneous action: the discomfort of men with acute myocardial infarction]. / Perda da espontaneidade da ação: o desconforto de homens que sofreram infarto agudo do miocárdio.

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 discomfort such as experienced by men who had suffered acute myocardial infarction (AMI). By resorting to the Symbolic Interactionism and to Grounded Theory methodology, data were collected by means of interviews with 13 men who had suffered AMI, at two health units in the city of São Paulo. The analysis led to building a theoretical model of such experience, made up by three phenomena and essentially prevaded by the discomfort of "undergoing loss of spontaneous action." This articles sums up the basic psychosocial process that emerges from that experience, and discusses its implications to question the clinical model of treatment, pointing to prevention as a further scope of action for nurses, besides raising issues to enhance nurse education.

[Selection of graduate nurses for admission to graduate programs in specialization areas]. / SELEÇÃO DE CANDIDATOS AOS CURSOS DE ESPECIALIZAÇÃO.

The authors analyse the selection process that is being used at the "Escota de Enfermagem da Universidade de São Paulo", for selection of graduate nurses the enrollment in the specialization program on Intensive Care Nursing. They refer the objective and the method used at the selection of candidates, as the results obtained.

[Contents of Revista da Escola de Enfermagem da USP: period of 1982 to 1986]. / ANÁLISE DA REVISTA DA ESCOLA DE ENFERMAGEM DA USP: PERÍODO DE 1982 a 1986.

To celebrate the 20th aniversary of the Revista da Escola de Enfermagem da USP, the authors decided to extend to the years of 1982 - 1986 the analysis of the first fifteen years of the periodical, above published, made by STEFANELLI et alii.

[Intolerance glucose control in the parenteral hyperalimentation]. / CONTROLE DA TOLERÂNCIA À GLICOSE NA NUTRIÇÃO PARENTERAL.

Ten non-diabetic surgical patients, submitted to parenteral hyperalimentation, were prospectively studied, in order to determine the incidence of glucose intolerance during this treatment. Additionally three methods for the control of glucose metabolism, namely glucosuria and gtycemia, given by reagent strips, and standard blood sugar, determined at the auto-analyser, were compared, with the purpose of establishing which of these tests is best suited for routine utilization. It was concluded that: 1) The incidence of glucose intolerance, defined as one or more episodes of positive glucosuria, or of blood sugar higher than 180mg/100 ml, was 60% in the first day of intravenous nutrition, and 80% if the whole period of treatment; 2) Glucose intolerance was more often detected by urine testing than by blood determinations, either with Dextrositx or at the a auto-analyser as the glucosuria test: was performed much more frequently; 3) It is concluded that although the findings of tests for glucosuria may not adequately reflect glucose metabolism in all circumstances, they represent the choice examination for basic monitoring of patients receiving intravenous hyperalimentation, as soon as renal function is satisfactory, because of its low expense and its easy application.