Mitochondria directly influence fertilisation outcome in the pig

The mitochondrion is explicitly involved in cytoplasmic regulation and is the cell's major generator of ATP. Our aim was to determine whether mitochondria alone could influence fertilisation outcome. In vitro, oocyte competence can be assessed through the presence of glucose-6-phosphate dehydrogenase (G6PD) as indicated by the dye, brilliant cresyl blue (BCB). Using porcine in vitro fertilisation (IVF), we have assessed oocyte maturation, cytoplasmic volume, fertilisation outcome, mitochondrial number as determined by mtDNA copy number, and whether mitochondria are uniformly distributed between blastomeres of each embryo. After staining with BCB, we observed a significant difference in cytoplasmic volume between BCB positive (BCB+) and BCB negative (BCB-) oocytes. There was also a significant difference in mtDNA copy number between fertilised and unfertilised oocytes and unequal mitochondrial segregation between blastomeres during early cleavage stages. Furthermore, we have supplemented BCB- oocytes with mitochondria from maternal relatives and observed a significant difference in fertilisation outcomes following both IVF and intracytoplasmic sperm injection (ICSI) between supplemented, sham-injected and non-treated BCB- oocytes. We have therefore demonstrated a relationship between oocyte maturity, cytoplasmic volume, and fertilisation outcome and mitochondrial content. These data suggest that mitochondrial number is important for fertilisation outcome and embryonic development. Furthermore, a mitochondrial pre-fertilisation threshold may ensure that, as mitochondria are diluted out during post-fertilisation cleavage, there are sufficient copies of mtDNA per blastomere to allow transmission of mtDNA to each cell of the post-implantation embryo after the initiation of mtDNA replication during the early postimplantation stages

Simulations of extensional flow in microrheometric devices

We present a detailed numerical study of the flow of a Newtonian fluid through microrheometric devices featuring a sudden contraction–expansion. This flow configuration is typically used to generate extensional deformations and high strain rates. The excess pressure drop resulting from the converging and diverging flow is an important dynamic measure to quantify if the device is intended to be used as a microfluidic extensional rheometer. To explore this idea, we examine the effect of the contraction length, aspect ratio and Reynolds number on the flow kinematics and resulting pressure field. Analysis of the computed velocity and pressure fields show that, for typical experimental conditions used in microfluidic devices, the steady flow is highly three-dimensional with open spiraling vortical structures in the stagnant corner regions. The numerical simulations of the local kinematics and global pressure drop are in good agreement with experimental results. The device aspect ratio is shown to have a strong impact on the flow and consequently on the excess pressure drop, which is quantified in terms of the dimensionless Couette and Bagley correction factors. We suggest an approach for calculating the Bagley correction which may be especially appropriate for planar microchannels

Nanoscaled discovery of a shunt rifamycin from Salinispora arenicola using a three-colour GFP-tagged Staphylococcus aureus macrophage infection assay

Antimicrobial resistance has emerged as a global public health threat, and development of novel therapeutics for treating infections caused by multi-drug resistant bacteria is urgent. Staphylococcus aureus is a major human and animal pathogen, responsible for high levels of morbidity and mortality worldwide. The intracellular survival of S. aureus in macrophages contributes to immune evasion, dissemination, and resilience to antibiotic treatment. Here, we present a confocal fluorescence imaging assay for monitoring macrophage infection by green fluorescent protein (GFP)-tagged S. aureus as a front-line tool to identify antibiotic leads. The assay was employed in combination with nanoscaled chemical analyses to facilitate the discovery of a new, active rifamycin analogue. Our findings indicate a promising new approach for the identification of antimicrobial compounds with macrophage intracellular activity. The antibiotic identified here may represent a useful addition to our armory in tackling the silent pandemic of antimicrobial resistance

Comprehensiveness and programmatic vulnerability to stds/hiv/aids in primary care

This study aimed to identify programmatic vulnerability to STDs/HIV/AIDS in primary health centers (PHCs). This is a descrip - tive and quantitative study carried out in the city of São Paulo. An online survey was applied (FormSUS platform), involving administrators from 442 PHCs in the city, with responses received from 328 of them (74.2%), of which 53.6% were nurses. At - tention was raised in relation to program - matic vulnerability in the PHCs regarding certain items of infrastructure, prevention, treatment, prenatal care and integration among services on STDs/HIV/AIDS care. It was concluded that in order to reach comprehensiveness of actions for HIV/ AIDS in primary health care, it is necessary to consider programmatic vulnerability, in addition to more investment and reor - ganization of services in a dialogue with the stakeholders (users, multidisciplinary teams, and managers, among others)

Knowledge and practices regarding child development among primary healthcare professionals

OBJECTIVE: To evaluate the knowledge and practices regarding child development among physicians working in primary healthcare units. METHOD: Cross-sectional descriptive study carried out at primary healthcare units in Embu, São Paulo, Brazil. Study procedures: 1) Evaluation of knowledge: test consisting of 20 multiple-choice questions on child development applied to all 31 physicians who were providing pediatric care at the primary healthcare units; 2) Evaluation of practices: semi-structured interview applied to a sample of 154 mothers/caregivers of children aged up to 36 months during follow-up visits at primary healthcare units in the municipality. For the comparisons of categorical variables (evaluation/advices about development in visits of children at different ages), the chi-square test was employed. RESULTS: The mean number of correct responses among physicians was 14.8. The error rate for seven questions was greater than 30% (sensory development, language acquisition, physiology of the nervous system, clinical and laboratory diagnosis of congenital infections and innate errors of metabolism) and the rate of correct responses was greater than 85% for four questions (motor and personal-social development markers, risk factors and genetic syndromes). Regarding practices, in 69 (45%) visits, the doctor asked the mother/caregiver's opinion about the child's development; in 80 (52%), the mother/caregiver said that the doctor assessed the development; and in 64 (42%), the mother/caregiver said that the doctor advised them on practices for child's stimulation. CONCLUSIONS: Faulty knowledge and practices regarding child development were identified among primary care professionals, indicating the need for continued education.OBJETIVO: Avaliar o conhecimento e as práticas sobre desenvolvimento infantil de médicos que atuam em Unidades Básicas de Saúde (UBS). MÉTODO: Estudo transversal, descritivo, realizado nas UBS de Embu (SP). Procedimentos do estudo: 1) avaliação do conhecimento por teste contendo 20 questões de múltipla escolha sobre desenvolvimento da criança aplicado a 31 médicos (universo) que prestam assistência pediátrica em UBS; 2) avaliação das práticas - entrevista semiestruturada aplicada para uma amostra de 154 mães/cuidadores que acompanhavam crianças com idade menor ou igual a 36 meses em consulta médica agendada em UBS do município. Para comparação de variáveis categóricas (avaliação/orientações sobre desenvolvimento em consultas de crianças de diferentes faixas etárias), utizou-se o qui-quadrado. RESULTADOS: A média de acertos dos médicos foi de 14,8 questões; sete questões apresentaram índices de erros superiores a 30% (desenvolvimento sensorial, aquisição de linguagem, fisiologia do sistema nervoso, diagnóstico clínico e laboratorial de infecções congênitas, erros inatos do metabolismo) e quatro questões apresentaram acertos acima de 85% (marcos do desenvolvimento motor, pessoal-social, fatores de risco e síndrome genética). Quanto às práticas, em 69 (45%) consultas o médico perguntou a opinião da mãe/cuidador sobre o desenvolvimento da criança, em 80 (52%) a mãe/cuidador referiu que o médico fez alguma pergunta e/ou avaliou o desenvolvimento e em 64 (42%) orientou sobre como estimular a criança. CONCLUSÕES: Identificaram-se falhas de conhecimento e nas práticas dos profissionais referentes ao desenvolvimento da criança, o que indica a necessidade de implementar educação permanente.UNIFESP Curso de MedicinaUNIFESP Projeto DesenvolverSecretaria Municipal de Saúde do EmbuUNIFESP Departamento de Pediatria Disciplina de Pediatria Geral e ComunitáriaUNIFESP, Curso de MedicinaUNIFESP, Projeto DesenvolverUNIFESP, Depto. de Pediatria Disciplina de Pediatria Geral e ComunitáriaSciEL

IT on teaching and learning process of visually impaired students

info:eu-repo/semantics/publishedVersio