Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo

Mesenchymal stem/stromal cells (MSCs) can differentiate into osteoblasts under appropriate conditions. PDGFRβ signaling controls MSC osteogenic potential both transcriptomically and in culture. Here, we present a “computer to the bench” protocol to analyze changes in MSC osteogenic potential at transcriptomic and cellular level in the absence of PDGFRβ. We detail the preparation of cells from mouse embryos, the analysis of transcriptomic changes from single-cell RNA-sequencing data, the procedure for MSC derivation and culture, and an osteogenic assay for functional validation. For complete details on the use and execution of this protocol, please refer to Sá da Bandeira et al. (2022).(1

Pollen morphology in tribe Dicomeae Panero and Funk (Asteraceae)

To better understand the taxonomy and phylogeny of the Dicomeae (Asteraceae) the pollen morphology of seven genera including 15 species of that tribe and six genera with seven species belonging to five related tribes was studied by use of light and scanning electron microscopy. The quantitative data were analysed by use of principal-components analysis (PCA). The exine ultrastructure of Erythrocephalum longifolium and Pleiotaxis rugosa was also studied by use of transmission electron microscopy. Three pollen types were distinguishable from the apertural, columellar, and spinular morphology and inter-spinular sculpture. A dichotomous key to these pollen types is proposed. The existence of the Dicomeae as an independent tribe and its subdivision in two subtribes (Dicominae and Pleiotaxinae) are supported by this study, which also suggests the Oldenburgieae and the Tarchonantheae are the closest tribes to Dicomeae

Cellular composition characterizing postnatal development and maturation of the mouse brain and spinal cord

The process of development, maturation, and regression in the central nervous system (CNS) are genetically programmed and influenced by environment. Hitherto, most research efforts have focused on either the early development of the CNS or the late changes associated with aging, whereas an important period corresponding to adolescence has been overlooked. In this study, we searched for age-dependent changes in the number of cells that compose the CNS (divided into isocortex, hippocampus, olfactory bulb, cerebellum, ‘rest of the brain’, and spinal cord) and the pituitary gland in 4–40-week-old C57BL6 mice, using the isotropic fractionator method in combination with neuronal nuclear protein as a marker for neuronal cells. We found that all CNS structures, except for the isocortex, increased in mass in the period of 4–15 weeks. Over the same period, the absolute number of neurons significantly increased in the olfactory bulb and cerebellum while non-neuronal cell numbers increased in the ‘rest of the brain’ and isocortex. Along with the gain in body length and weight, the pituitary gland also increased in mass and cell number, the latter correlating well with changes of the brain and spinal cord mass. The majority of the age-dependent alterations (e.g., somatic parameters, relative brain mass, number of pituitary cells, and cellular composition of the cerebellum, isocortex, rest of the brain, and spinal cord) occur rapidly between the 4th and 11th postnatal weeks. This period includes murine adolescence, underscoring the significance of this stage in the postnatal development of the mouse CNS

Accidents and violence in childhood: survey evidence of emergency care for external causes - Brazil, 2009

Compreender as características e a magnitude das causas externas (acidentes e violência) em crianças de 0 a 9 anos de idade torna-se cada vez mais importante em Saúde Pública. O objetivo do presente artigo foi analisar os atendimentos de emergência por causas externas em crianças. Utilizaram-se dados do Sistema de Vigilância de Violências e Acidentes em Serviços Sentinelas de Urgência e Emergência (Inquérito VIVA), realizado em 74 serviços de urgência do Distrito Federal e 23 capitais no ano 2009. Analisaram-se dados de 7.123 crianças: 6.897 (96,7%) vítimas de acidentes e 226 (3,3%) de violência. Em comparação às vítimas de violência, os atendimentos por acidentes foram mais frequentes entre crianças de 2 a 5 anos, de pele branca e ocorridos no domicílio (p < 0,05). Dentre os acidentes, as quedas e queimaduras predominaram no grupo de 0 a 1 ano, enquanto os acidentes de transporte foram mais frequentes no grupo de 6 a 9 anos (p < 0,001). Quanto às violências, atendimentos por negligência e agressão física predominaram, respectivamente, nos grupos extremos de faixa etária, sendo um familiar identificado como agressor (p < 0,001). Informações sobre ocorrência de causas externas em crianças podem apoiar políticas de promoção da saúde, além de orientar profissionais de saúde, educadores e famílias na prevenção destas causas.Understanding the characteristics and magnitude of accidents and violence due to external causes in children from 0 to 9 years of age is becoming ever more important in Public Health. The scope of this paper was to analyze emergency care for accidents due to external causes in children. The Sentinel Urgency and Emergency Services of the Violence and Accident Vigilance System (VIVA Survey), performed in 74 urgency services in the Federal District and 23 State capitals in 2009 was used. Data of 7,123 children were analyzed: 6,897 (96.7%) victims of accidents and 226 (3.3%) of violence. In comparison with victims of violence, the visits for accidents were more frequent among white children from 2 to 5 years old occurring in the home (p &lt; 0.05). Among the accidents, falls and burns predominated in the 0 to 1group, while traffic accidents were most frequent in the 6 to 9-year-old age group (p < 0.001). With respect to violence, the visits for neglect and physical assault predominated, respectively, in extreme age groups, with someone in the family being the perpetrator (p < 0.001). Information on the occurrence of external causes in children may support health promotion policies, besides guiding health professionals, teachers and families in the prevention of such causes

Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700