Tamoxifen-Induced Cre-loxP Recombination Is Prolonged in Pancreatic Islets of Adult Mice

Tamoxifen (Tm)-inducible Cre recombinases are widely used to perform gene inactivation and lineage tracing studies in mice. Although the efficiency of inducible Cre-loxP recombination can be easily evaluated with reporter strains, the precise length of time that Tm induces nuclear translocation of CreERTm and subsequent recombination of a target allele is not well defined, and difficult to assess. To better understand the timeline of Tm activity in vivo, we developed a bioassay in which pancreatic islets with a Tm-inducible reporter (from Pdx1PB-CreERTm;R26RlacZ mice) were transplanted beneath the renal capsule of adult mice previously treated with three doses of 1 mg Tm, 8 mg Tm, or corn oil vehicle. Surprisingly, recombination in islet grafts, as assessed by expression of the β-galactosidase (β-gal) reporter, was observed days or weeks after Tm treatment, in a dose-dependent manner. Substantial recombination occurred in islet grafts long after administration of 3×8 mg Tm: in grafts transplanted 48 hours after the last Tm injection, 77.9±0.4% of β-cells were β-gal+; in β-cells placed after 1 week, 46.2±5.0% were β-gal+; after 2 weeks, 26.3±7.0% were β-gal+; and after 4 weeks, 1.9±0.9% were β-gal+. Islet grafts from mice given 3×1 mg Tm showed lower, but notable, recombination 48 hours (4.9±1.7%) and 1 week (4.5±1.9%) after Tm administration. These results show that Tm doses commonly used to induce Cre-loxP recombination may continue to label significant numbers of cells for weeks after Tm treatment, possibly confounding the interpretation of time-sensitive studies using Tm-dependent models. Therefore, investigators developing experimental approaches using Tm-inducible systems should consider both maximal recombination efficiency and the length of time that Tm-induced Cre-loxP recombination occurs

Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture

The human cerebral cortex develops through an elaborate succession of cellular events that, when disrupted, can lead to neuropsychiatric disease. The ability to reprogram somatic cells into pluripotent cells that can be differentiated in vitro provides a unique opportunity to study normal and abnormal corticogenesis. Here, we present a simple and reproducible 3D culture approach for generating a laminated cerebral cortex–like structure, named human cortical spheroids (hCSs), from pluripotent stem cells. hCSs contain neurons from both deep and superficial cortical layers and map transcriptionally to in vivo fetal development. These neurons are electrophysiologically mature, display spontaneous activity, are surrounded by nonreactive astrocytes and form functional synapses. Experiments in acute hCS slices demonstrate that cortical neurons participate in network activity and produce complex synaptic events. These 3D cultures should allow a detailed interrogation of human cortical development, function and disease, and may prove a versatile platform for generating other neuronal and glial subtypes in vitro

The performance of various anthropometric assessment methods for predicting low birth weight in pregnant women Desempenho de diferentes métodos de avaliação antropométrica de gestantes na predição de baixo peso ao nascer

OBJECTIVES: to assess the performance of various anthropometric methods for the evaulation of the nutritional status of pregnant women as a means of predicting low birth weight (LBW). METHODS: a descriptive cross-cutting study carried out among 433 pregnant women (>20 years) attending a Public Maternity Hospital in Rio de Janeiro, Brazil. The adequacy of the weight gain at the end of the pregnancy was evaluated in accordance with the proposals of the Institute of Medicine and the Brazilian Ministry of Health. The sensitivity, specificity and accuracy of the adequacy of weight gain at the end of the pregnancy or nutritional state of mother as a predictor of low birth weight were calculated. RESULTS: the sensitivity of the various methods varied from 63.1% to 68.4% and the specificity from 71.2% to 75.1%. The adapted Institute of Medicine proposal drawn up by the Brazilian Ministry of Health, according to the classification of the pre-delivery nutritional status of the mother according to the World Health Organization cutoff points showed itself to be the most accurate (74.5%), this being the most adequate method for nutritional triage for reason of its association with low birth weight (OR=4.10; 95%CI=1.53-10.92). CONCLUSIONS: the best proposals for this population are those of the Institute of Medicine and the Brazilian Ministry of Health. Further studies aiming to ascertain the most appropriate methods of anthropometric evaluation for different populations should be encouraged.<br>OBJETIVOS: avaliar o desempenho de diferentes métodos antropométricos para avaliação nutricional de gestantes para predizer o baixo peso ao nascer (BPN). MÉTODOS: estudo descritivo do tipo transversal, realizado com 433 puérperas (>20 anos) atendidas numa Maternidade Pública do Rio de Janeiro, Brasil. A adequação do ganho de peso ao final da gestação foi avaliada segundo as propostas do Institute of Medicine e do Ministério da Saúde. Calculou-se a sensibilidade, a especificidade e a acurácia das variáveis adequação do ganho de peso gestacional total ou adequação do estado nutricional materno ao final da gestação na predição do BPN. RESULTADOS: a sensibilidade dos métodos variou de 63,1% a 68,4% e a especificidade de 71,2% a 75,1%. A adaptação da proposta do Institute of Medicine elaborada pelo Ministério da Saúde, segundo a classificação do estado nutricional pré-gestacional pelos pontos de corte da Organização Mundial da Saúde apresentou maior acurácia (74,5%), sendo este último o mais adequado para triagem nutricional pela sua associação com o BPN (OR=4,10; IC95%=1,53-10,92). CONCLUSÕES: os melhores para esta população foram as propostas do Institute of Medicine e do Ministério da Saúde. Estudos visando reconhecer os métodos de avaliação antropométrica mais adequados na gestação devem ser estimulados em diferentes populações