Why not "do simple things in a simple way": Use of the Pap test as the first step in screening genetic stability for human cultured stem cell therapy?

The aim of this study was to analyze adipose tissue-derived mesenchymal stem cells (AT-MSCs) using the Pap test as a first screening step to evaluate genetic stability. Human adipose tissue from six healthy female donors was obtained from elective liposuction procedures. The cells were isolated, cultivated at P2/P3, characterized by flow cytometric analysis, and differentiation induced. The AT-MSCs were stained by Papanicolaou staining and analyzed according to the Bethesda classification, and viability-apoptosis relationships were evaluated. The results of the Pap test for Sample I indicated high-grade alterations consistent with genetic instability; for Samples II-V, atypical cells of undetermined significance; and for Sample VI, normal cells. These results demonstrate the potential of using the Pap test as an initial screening step to evaluate the genetic stability of cultured AT-MSCs and also suggest its use for other adherent cells such as embryonic stem cells or induced pluripotent stem cells

Production and characterization of protease from Penicillium aurantiogriseum URM 4622

Proteases with new properties are required due to their increasing industrial importance. In this work, the optimal fermentation conditions for the production of a protease from Penicillium aurantiogriseum dierchx (URM-4622) are presented together with partial characterization of the protease catalytic properties. The batch fermentation conditions that allow for the highest specific proteolytic activity are 26 ºC, pH 7.0, and 25 % saturation dissolved O2 concentration. The obtained protease is stable over a wide range of pH (5.8 to 9.5) and temperature (25 to 40 ºC) values. In the presence of Zn2+ a 26 % reduction in the enzyme proteolytic activity occurs and, in contrast, Mn2+ enhances its activity by 28.9 %. 96.2 % and 70.8 % of the protease activity are maintained after 90 min incubation in 5 and 10 % (v/v) H2O2 aqueous solutions, respectively. PMSF inhibition reveals that this enzyme is a serine protease. Protease is able to hydrolyze different proteins

Expectativa do resultado pós-operatório pela ótica do idoso informatizado com síndrome do olho seco e catarata - ética e evidência

Introdução: Explorar a relação do oftalmologista com paciente idoso e intelectualmente diferenciado, do ponto de vista científico, sobre síndrome de olho seco e resultados de cirurgia de catarata. Relato do caso: Médico, pesquisador, branco, 74,5 anos, portador de catarata e olho seco, ansioso por ter obtido informações, em bases de dados, sobre riscos do aumento dos sintomas da síndrome de olho seco no pós-operatório da facectomia. Foi adequadamente avaliado pelo especialista, assim se candidatando ao procedimento de forma livre e espontânea. Comentários: As informações técnicas pelo profissional especializado tiveram seu cerne em medicina baseada em vidência e ética, dando suporte para redução da ansiedade do paciente. Há necessidade do preparo do especialista para lidar com os pacientes idosos do novo milênio, atualizados na era cibernética

The Barretos Cancer Hospital Animal Facility: implementation and results of a dedicated platform for preclinical oncology models

The Barretos Cancer Hospital Animal Facility (BCHAF) is a unique facility in Brazil exclusively dedicated to working with animal models for cancer research. In this article, we briefly present our modern facility and the main experiments performed, focusing on mutant strains of mice (PTCH-knockout and ApcMin mice), xenograft models, and patient-derived xenografts (PDXs). Our results show the progress and challenges in establishing these models and the need for having an appropriate representation of our cancer population to better understand tumor biology and to identify cancer biomarkers, which could be putatively targeted, allowing for personalized therapy.This study was funded by the Public Ministry of Labor Campinas (Research, Prevention and Education of Occupational Cancer) and by Pio XII Foundation, Barretos Cancer Hospital internal funds, Grant Number: 13/2021

Clonidine-stimulated growth hormone concentrations (cut-off values) measured by immunochemiluminescent assay (ICMA) in children and adolescents with short stature

OBJECTIVES: To establish cut-off values for growth hormone concentrations using clonidine as a secretagogue and an immunochemiluminescent assay as the method of measurement and to analyze the response time as well as the influence of gender, nutritional status and pubertal stage. METHODS: A total of 225 tests were performed in 3 patient groups, categorized as group 1 (normal), group 2 (idiopathic short stature) and group 3 (growth hormone deficiency). Among the 199 disease-free individuals, 138 were prepubertal, and 61 were pubertal. Clonidine (0.1 mg/m2) was orally administered, and the growth hormone level was measured by immunochemiluminescent assay. The growth hormone peak and the difference between the growth hormone peak and the baseline level were then analyzed. Statistical analyses were performed using Student’s t-test or the Mann-Whitney test and Kruskal-Wallis test followed by Dunn’s post hoc test. Cut-off values were determined using a receiver operating characteristic curve. RESULTS: Group 1 and group 2 had no difference in growth hormone peak, gender, body mass index standard deviation score, or pubertal stage. Group 3 exhibited a significantly lower growth hormone peak than the other groups did. The receiver operating characteristic curve demonstrated that growth hormone concentrations ≥ 3.0 ng/mL defined responsiveness to clonidine. In total, 3.02% of individuals in group 1 and group 2 were considered false positive, i.e., these children lacked growth hormone deficiency and had a peak below 3.0 ng/mL. CONCLUSION: Clonidine-stimulated growth hormone concentrations ≥3 ng/mL, as measured by immunochemiluminescent assay, suggest responsiveness to the stimulus regardless of gender, body mass index standard deviation score or pubertal stage

Sequência rápida de intubação na emergência - paciente suspeito de COVID19

Introdução: O COVID-19 possui como uma das principais características o acometimento pulmonar, podendo gerar quadros de pneumonias virais em pacientes graves, que poderão necessitar de intubação orotraqueal para a estabilidade clínica do seu quadro. Contudo durante o procedimento para acesso das vias aéreas desses pacientes podem ser gerados aerossóis, colocando em risco a saúde dos profissionais de saúde que realizam o procedimento. Objetivo: Revisar as principais publicações realizadas até o presente momento sobre a sequência rápida de intubação em pacientes suspeito de COVID-19, descrevendo as melhores evidências para a execução deste procedimento de forma segura. Metodologia: O presente estudo constitui-se de uma revisão de literatura realizada com artigos publicados em 2020 e com o Protocolo para intubação orotraqueal em pacientes suspeitos de COVID-19 realizado pela Associação Brasileira de Medicina de Emergência em conjunto com Associação de Medicina Intensiva Brasileira. As bases de dados utilizadas foram: PubMed e Google Scholar. As palavras-chave utilizadas na busca foram: ‘Sequência Rápida de Intubação”, “Intubação”, “COVID-19”, “SARCOV-2”, “Intubation”. Resultados e Discussão: Dos pacientes infectados pelo novo coronavírus, 5 a 26% necessitam de internação em Unidade de terapia intensiva para tratamento respiratório. Dentre esses, cerca de 3,2% dos pacientes infectados precisaram de intubação e ventilação invasiva durante a doença. Para que seja realizada a intubação orotraqueal são necessários Equipamento de Proteção Individual (EPIs). Todo material de intubação é preparado fora da área de risco de contaminação e o ventilador mecânico programado com os parâmetros iniciais da ventilação. A utilização de EPIs e seguir o protocolo de proteção são medidas de extrema importância. Na China, estima-se que cerca de 3,5% dos profissionais de saúde foram infectados com COVID-19, e sua taxa de mortalidade foi de cerca de 0,3%. O preconizado é a utilização da sequência rápida de intubação. A utilização de videolaringoscópios propicia  maior segurança, possibilitando que o intubador fique mais afastado da via aérea, sem comprometer a visualização. Durante o procedimento é  importante que se conecte o sistema de aspiração fechado ao respirador e que faça uma pré-oxigenação no paciente. Se possível, evitar a ventilação assistida devido a produção de aerossol. Após a intubação, a confirmação da localização do tubo deve ser feita através da capnografia, evitando o uso do dispositivo Bolsa-Válvula-Máscara e estetoscópio pelos riscos de contaminação e pela menor eficácia. Os fármacos utilizados incluem Lidocaína que vai atuar inibindo os reflexos laríngeos e dando uma maior potência de ação aos outros fármacos. Cetamina, anestésico dissociativo,que apresenta ações broncodilatadoras com grande  estabilidade hemodinâmica. Succinilcolina ou Rocurônio, bloqueadores neuromusculares que relaxam a musculatura esquelética e facilitam a intubação, evitando também que o paciente tenha tosse durante o processo. Midazolam e Fentanil são utilizados para sedação e analgesia logo após a intubação. Conclusão: A sequencia rápida de intubação é cada vez mais realizada nas emergências devido a atual pandemia do COVID19. No intuito de diminuir a contaminação dos profissionais de saúde protocolos foram criados. O uso de EPIs, videolaringoscópio e a não ventilação fazem parte das medidas preconizadas.&nbsp

The impact of demographic and clinical characteristics on diabetic painful neuropathy

Introduction: Diabetic neuropathy (DN) is one of the most devastating complications of diabetes mellitus; however, in contrast to other countries, there are no scientific studies in Portugal evaluating the impact of demographic and clinical characteristics of this pathological entity. The aim of this study was to evaluate the impact of gender, metabolic control, age of diabetic patients, as well as time of disease progression, the appearance of complaints related to neuropathic pain. Material and methods: A multicentre study with a non-probabilistic, convenience sample of 359 patients was performed employing the quantitative method, using the Statistical Package for Social Science 24 software. The p-value of p < 0.05 was defined to consider a result statistically significant. The Spearman correlation coefficient (r) was determined to determine the relationship between categorical variables. Results: There was no statistically significant difference in the prevalence of DN between genders (p = 0.633 and r = 0.025). There was a statistically significant relationship between the value of HbA1c and DN, with p = 0.010 and r = 0.136. There is a relationship between age and complaints of neuropathic pain, with p = 0.034 and r = 0.112. The variable, time of disease progression, is also correlated with the appearance of complaints of neuropathic pain with p = 0.020 and r = 0.112. Conclusion: The prevalence of neuropathic pain in subjects with diabetes is not negligible and is associated with modifiable risk factors that can be identified, possibly modified and prevented. The correct approach for these patients, which involves screening and early treatment, is decisive improving functionality and quality of life.info:eu-repo/semantics/publishedVersio

Production and characterization of protease from Penicillium aurantiogriseum URM 4622

Abstract Proteases with new properties are required due to their increasing industrial importance. In this work, the optimal fermentation conditions for the production of a protease from Penicillium aurantiogriseum dierchx (URM-4622) are presented together with partial characterization of the protease catalytic properties. The batch fermentation conditions that allow for the highest specific proteolytic activity are 26 ºC, pH 7.0, and 25 % saturation dissolved O 2 concentration. The obtained protease is stable over a wide range of pH (5.8 to 9.5) and temperature (25 to 40 ºC) values. In the presence of Zn 2+ a 26 % reduction in the enzyme proteolytic activity occurs and, in contrast, Mn 2+ enhances its activity by 28.9 %. 96.2 % and 70.8 % of the protease activity are maintained after 90 min incubation in 5 and 10 % (v/v) H 2 O 2 aqueous solutions, respectively. PMSF inhibition reveals that this enzyme is a serine protease. Protease is able to hydrolyze different proteins