Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson). This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use

The Relationship Between Bleeding on Probing and Subgingival Deposits. An Endoscopical Evaluation

none4Background: Bleeding on probing (BOP) is an indicator of tissue inflammatory response to bacterial pathogens. Because anatomical limitations the entity and physical state of microbial aggregations located under the gingival margin and their relations to BOP have been hardly investigated till now. The recent introduction of the endoscopy has allowed clinicians to view the subgingival environment in a non-traumatic way. Aim of this study is to evaluate the correlation between BOP and subgingival deposits by using this new technology. Methods: At one-month revaluation of 16 periodontal patients treated with scaling and root planning, 107 teeth (642 individual sites) were evaluated for plaque index (PI), gingival index (GI), probing pocket depth (PPD), bleeding on probing (BOP), endoscopic biofilm index (EBI) and endoscopic calculus index (ECI). Results: A linear association between BOP and PD, EBI, and ECI was detected. The BOP provided a high level of specificity but quite low sensitivity values both for ECI (sensitivity 40%, specificity 86%) and EBI (sensitivity 37%, specificity 89%). The BOP sensitivity was directly linked to the amount of subgingival deposits. Conclusions: This study demonstrates a direct relationship between BOP and presence/amount of subgingival deposits. More investigations on larger samples are however needed.noneChecchi l.; Montevecchi M.; Checchi V.; Zappulla F.Checchi l.; Montevecchi M.; Checchi V.; Zappulla F

Capturing the cloud of diversity reveals complexity and heterogeneity of MRSA carriage, infection and transmission.

Genome sequencing is revolutionizing clinical microbiology and our understanding of infectious diseases. Previous studies have largely relied on the sequencing of a single isolate from each individual. However, it is not clear what degree of bacterial diversity exists within, and is transmitted between individuals. Understanding this 'cloud of diversity' is key to accurate identification of transmission pathways. Here, we report the deep sequencing of methicillin-resistant Staphylococcus aureus among staff and animal patients involved in a transmission network at a veterinary hospital. We demonstrate considerable within-host diversity and that within-host diversity may rise and fall over time. Isolates from invasive disease contained multiple mutations in the same genes, including inactivation of a global regulator of virulence and changes in phage copy number. This study highlights the need for sequencing of multiple isolates from individuals to gain an accurate picture of transmission networks and to further understand the basis of pathogenesis.Thanks to Dr Alex O’Neill, University of Leeds and Dr Matthew Ellington, Public Health England for provision of RN4220 and RN4200mutS. We thank the core sequencing and informatics team at the Wellcome Trust Sanger Institute for sequencing of the isolates described in this study. This work was supported by a Medical Research Council Partnership grant (G1001787/1) held between the Department of Veterinary Medicine, University of Cambridge (M.A.H.), the School of Clinical Medicine, University of Cambridge (S.J.P.), the Moredun Research Institute, and the Wellcome Trust Sanger Institute (J.P. and S.J.P). S.J.P. receives support from the NIHR Cambridge Biomedical Research Centre. M.T.G.H., S.R.H. and J.P. were funded by Wellcome Trust grant no. 098051. G.G.R.M. was funded by an MRC studentship.This is the final version of the article. It first appeared from Nature Publishing Group via http://dx.doi.org/10.1038/ncomms756

Perception of COVID-19 pandemic restrictions on dental researchers

BACKGROUND/OBJECTIVES: Historical evidence shows a gender‐based disproportionate effect of pandemics across different populations. In 2020, the coronavirus disease 2019 (COVID‐19) pandemic began spreading its devastating effects worldwide. The goal of the present study was to investigate the effect of the COVID‐19 pandemic on research productivity, work‐life arrangements, and mental health of dental professionals worldwide with focus on gender differences. METHODS: A 38‐item survey, concerning demographics, career stage, employer support, family structure, mental health, and relationships, was distributed to 7692 active members of the International Association for Dental Research. Bivariate associations between independent variables and the primary outcome variable were tested using Spearman's correlation test. A logistic regression model was used to assess the simultaneous, independent associations between each variable and researcher productivity. RESULTS: A total of 722 responses were obtained, indicating a 9.4% response rate. Higher productivity was reported by male respondents (p = 0.021), and by those in senior career stages (p = 0.001). Institutional support was associated with higher productivity (p < 0.0001). Lower productivity was reported by younger researchers (p = 0.003). Remote work negatively affected productivity (p < 0.0001) and female respondents reported working more hours, regardless of work location (p = 0.004). Poor mental health was associated with low productivity (p < 0.0001). CONCLUSIONS: Our results showed that the COVID‐19 pandemic significantly affected dental professionals’ perceived productivity and mental health around the globe. Younger individuals and women were disproportionally affected, and institutional support had a significant influence to mitigate effects of the pandemic for dental researchers

Internal consistency and reliability of Primary Care Assessment Tool (PCATool-Brasil) for child health services

Health strategies based on primary health care have been expanding in Brazil. An instrument applied to users, the Primary Care Assessment Tool (the PCATool), which measures the extent of primary care, has been validated in the United States. We sought to adapt the PCATool to Brazil and analyze its validation and reliability through a cross-sectional validation study of the Child PCATool.Validation included: translation, back-translation, adaptation, debriefing, content and construct validate, internal consistency, and reliability analysis. The questionnaire was applied to 468 parents or guardians of children registered with 18 primary health services in Porto Alegre, representing the services’ normal pediatric population. Using factor analysis, 8 domains were identified, with Cronbach’s α ranging from 0.74 to 0.88.Validation resulted in a 45-item scale, divided into 8 attributes (Access to First Contact, Continuity, Coordination, 3 attributes of Comprehensiveness, Community Orientation, and Family Orientation). These results showed that the PCATool-Brasil has adequate validity and reliability and could be used as a national instrument to evaluate primary health care after its application to other population settings in the country.Fundação de Amparo à Pesquisa, Estado do Rio Grande do Sul e Escola de Saúde Pública, Rio Grande do Su