A Magnetic Field-Based Wearable Respiration Sensor for Real-Time Monitoring During Pulmonary Rehabilitation.

OBJECTIVE: This work explores Hall effect sensing paired with a permanent magnet, in the context of pulmonary rehabilitation exercise training. METHODS: Experimental evaluation was performed considering as reference the gold-standard of respiratory monitoring, an airflow transducer, and performance was compared to another wearable device with analogous usability - a piezoelectric sensor. A total of 16 healthy participants performed 15 activities, representative of pulmonary rehabilitation exercises, simultaneously using all devices. Evaluation was performed based on detection of flow reversal events and key respiratory parameters. RESULTS: Overall, the proposed sensor outperformed the piezoelectric sensor with a mean ratio, precision, and recall of 0.97, 0.97, and 0.95, respectively, against 0.98, 0.90, and 0.88. Evaluation regarding the respiratory parameters indicates an adequate accuracy when it comes to breath cycle, inspiration, and expiration times, with mean relative errors around 4% for breath cycle and 8% for inspiration/expiration times, despite some variability. Bland-Altman analysis indicates no systematic biases. CONCLUSION: Characterization of the proposed sensor shows adequate monitoring capabilities for exercises that do not rely heavily on torso mobility, but may present a limitation when it comes to activities such as side stretches. SIGNIFICANCE: This work provides a comprehensive characterization of a magnetic field-based respiration sensor, including a discussion on its robustness to different algorithm thresholds. It proves the viability of the sensor in a range of exercises, expanding the applicability of Hall effect sensors as a feasible wearable approach to real-time respiratory monitoring.

Circulating Blood Lymphocytes and Acute Pancreatitis Severity: A Systematic Review.

Acute pancreatitis is an acute inflammatory process of the pancreas with high prevalence and varying degrees of severity that can be potentially life-threatening. Much is still unknown about which mechanisms determine the course and severity of acute pancreatitis. The primary objective of this review is to identify the potential association between circulating lymphocytes and the severity of acute pancreatitis. A systematic search was performed in Medline, Web of Science, Cochrane Central Register of Controlled Trials and ClinicalTrails.gov. The authors independently did the selection process as well as data extraction that was recorded into a flow diagram following the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols (PRISMA-P). Our initial search identified 27,783 studies which were narrowed down to 13 by applying strict inclusion and exclusion algorithms. The consistent findings across the studies indicated that peripheral blood lymphocytes are related to acute pancreatitis severity.

PreEpiSeizures: description and outcomes of physiological data acquisition using wearable devices during video-EEG monitoring in people with epilepsy.

The PreEpiSeizures project was created to better understand epilepsy and seizures through wearable technologies. The motivation was to capture physiological information related to epileptic seizures, besides Electroencephalography (EEG) during video-EEG monitorings. If other physiological signals have reliable information of epileptic seizures, unobtrusive wearable technology could be used to monitor epilepsy in daily life. The development of wearable solutions for epilepsy is limited by the nonexistence of datasets which could validate these solutions. Three different form factors were developed and deployed, and the signal quality was assessed for all acquired biosignals. The wearable data acquisition was performed during the video-EEG of patients with epilepsy. The results achieved so far include 59 patients from 2 hospitals totaling 2,721 h of wearable data and 348 seizures. Besides the wearable data, the Electrocardiogram of the hospital is also useable, totalling 5,838 h of hospital data. The quality ECG signals collected with the proposed wearable is equated with the hospital system, and all other biosignals also achieved state-of-the-art quality. During the data acquisition, 18 challenges were identified, and are presented alongside their possible solutions. Though this is an ongoing work, there were many lessons learned which could help to predict possible problems in wearable data collections and also contribute to the epilepsy community with new physiological information. This work contributes with original wearable data and results relevant to epilepsy research, and discusses relevant challenges that impact wearable health monitoring.

Bioactive cellulose acetate nanofiber loaded with annatto support skeletal muscle cell attachment and proliferation.

Electrospinning emerged as a promising technique to produce scaffolds for cultivated meat in function of its simplicity, versatility, cost-effectiveness, and scalability. Cellulose acetate (CA) is a biocompatible and low-cost material that support cell adhesion and proliferation. Here we investigated CA nanofibers, associated or not with a bioactive annatto extract (CA@A), a food-dye, as potential scaffolds for cultivated meat and muscle tissue engineering. The obtained CA nanofibers were evaluated concerning its physicochemical, morphological, mechanical and biological traits. UV-vis spectroscopy and contact angle measurements confirmed the annatto extract incorporation into the CA nanofibers and the surface wettability of both scaffolds, respectively. SEM images revealed that the scaffolds are porous, containing fibers with no specific alignment. Compared with the pure CA nanofibers, CA@A nanofibers showed increased fiber diameter (420 ± 212 nm vs. 284 ± 130 nm). Mechanical properties revealed that the annatto extract induces a reduction of the stiffness of the scaffold. Molecular analyses revealed that while CA scaffold favored C2C12 myoblast differentiation, the annatto-loaded CA scaffold favored a proliferative state of these cells. These results suggest that the combination of cellulose acetate fibers loaded with annatto extract may be an interesting economical alternative for support long-term muscle cells culture with potential application as scaffold for cultivated meat and muscle tissue engineering.

aPKC regulates apical constriction to prevent tissue rupture in the Drosophila follicular epithelium.

Apical-basal polarity is an essential epithelial trait controlled by the evolutionarily conserved PAR-aPKC polarity network. Dysregulation of polarity proteins disrupts tissue organization during development and in disease, but the underlying mechanisms are unclear due to the broad implications of polarity loss. Here, we uncover how Drosophila aPKC maintains epithelial architecture by directly observing tissue disorganization after fast optogenetic inactivation in living adult flies and ovaries cultured ex vivo. We show that fast aPKC perturbation in the proliferative follicular epithelium produces large epithelial gaps that result from increased apical constriction, rather than loss of apical-basal polarity. Accordingly, we can modulate the incidence of epithelial gaps by increasing and decreasing actomyosin-driven contractility. We traced the origin of these large epithelial gaps to tissue rupture next to dividing cells. Live imaging shows that aPKC perturbation induces apical constriction in non-mitotic cells within minutes, producing pulling forces that ultimately detach dividing and neighboring cells. We further demonstrate that epithelial rupture requires a global increase of apical constriction, as it is prevented by the presence of non-constricting cells. Conversely, a global induction of apical tension through light-induced recruitment of RhoGEF2 to the apical side is sufficient to produce tissue rupture. Hence, our work reveals that the roles of aPKC in polarity and actomyosin regulation are separable and provides the first in vivo evidence that excessive tissue stress can break the epithelial barrier during proliferation.

EpiBOX: An Automated Platform for Long-Term Biosignal Collection.

Biosignals represent a first-line source of information to understand the behavior and state of human biological systems, often used in machine learning problems. However, the development of healthcare-related algorithms that are both personalized and robust requires the collection of large volumes of data to capture representative instances of all possible states. While the rise of flexible biosignal acquisition solutions has enabled the expedition of data collection, they often require complicated frameworks or do not provide the customization required in some research contexts. As such, EpiBOX was developed as an open-source, standalone, and automated platform that enables the long-term acquisition of biosignals, passable to be operated by individuals with low technological proficiency. In particular, in this paper, we present an in-depth explanation of the framework, methods for the evaluation of its performance, and the corresponding findings regarding the perspective of the end-user. The impact of the network connection on data transfer latency was studied, demonstrating innocuous latency values for reasonable signal strengths and manageable latency values even when the connection was unstable. Moreover, performance profiling of the EpiBOX user interface (mobile application) indicates a suitable performance in all aspects, providing an encouraging outlook on adherence to the system. Finally, the experience of our research group is described as a use case, indicating a promising outlook regarding the use of the EpiBOX framework within similar contexts. As a byproduct of these features, our hope is that by empowering physicians, technicians, and monitored subjects to supervise the biosignal collection process, we enable researchers to scale biosignal collection.

Confidencialidade em medicina ocupacional: protegendo informações / Confidentiality in occupational medicine: protecting information / Confidencialidad en medicina del trabajo: protección de información

Resumo A confidencialidade é elemento central da relação médico-paciente e está associada à boa qualidade do atendimento. Contudo, pode ser rompida em conformidade com as normas éticas e legais estabelecidas no país. Este estudo objetiva mostrar os principais aspectos da confidencialidade em medicina ocupacional. Para isso, realizou-se revisão narrativa de literatura sobre o tema, utilizando bases de dados de livre acesso e embasando-se nos códigos de ética médica. A atuação do médico do trabalho envolve o trabalhador, outros profissionais não médicos e o empregador, situação capaz de desencadear conflitos, requerendo que o médico conheça suas obrigações e limites ético-legais. A proteção da confidencialidade respeita os direitos humanos, mas dilemas podem surgir, não bastando obedecer aos ditames éticos, mas sendo necessário essencialmente seguir as normas legais. Este estudo busca mostrar os principais aspectos éticos e legais atualizados referentes à saúde ocupacional.

Abstract Confidentiality is a central element of the physician-patient relationship and is associated with good quality of care. However, it may be broken in accordance with the ethical and legal standards established in the country. This study aims to show the main aspects of confidentiality in occupational medicine. For this, a narrative review of the literature on the subject was carried out, using free access databases and based on the codes of medical ethics. The occupational physician's performance involves the worker, other non-medical professionals and the employer, a situation that may trigger conflicts, requiring physicians to know their obligations and ethical-legal limits. The protection of confidentiality respects human rights, but dilemmas may arise, not only to obey ethical precepts, but to follow legal norms. This study seeks to show the main and updated ethical and legal aspects regarding occupational health.

Resumen La confidencialidad es clave en la relación médico-paciente y está asociada a buena calidad de la atención. Sin embargo, está sujeta a una quiebra de conformidad a lo establecido en las normas éticas y legales en el país. Este estudio pretende mostrar los principales aspectos de confidencialidad en la medicina del trabajo. Para ello, se realizó una revisión narrativa de la literatura en las bases de datos de acceso abierto basándose en códigos de ética médica. El actuar del médico del trabajo involucra al trabajador, a profesionales no médicos y al empleador, lo que puede desencadenar conflictos requiriendo que el médico conozca sus obligaciones y límites ético-legales. La protección de la confidencialidad respeta los derechos humanos, pero pueden surgir dilemas y no solo bastará atenerse a los dictámenes éticos, sino seguir fundamentalmente las normas legales. Los resultados mostraron los principales aspectos éticos y legales actualizados relacionados con la salud laboral.

Perfil de mulheres climatéricas em Estratégia de Saúde da Família no interior paulista / Health profile of climacteric women from the Family Health Strategy in the countryside of the state of São Paulo

Introdução: o climatério pode ser influenciado por mudanças psicossociais simultâneas, que associadas às alterações hormonais, como o hipoestrogenismo e a diminuição dos níveis de progesterona, podem intensificar os sintomas e agravos na mulher. Objetivo: identificar os perfis sociodemográficos, obstétricos, ginecológicos, de saúde e hábitos de vida das mulheres climatéricas atendidas na rede básica de saúde, por meio da aplicação de questionário, escalas de Hamilton de Ansiedade e Depressão, e Índice Menopausal de Kupperman e Blatt. Material e Método: estudo transversal descritivo exploratório de relato de experiência, desenvolvido por graduandas em Medicina e Enfermagem. As participantes apresentam idade entre 45 e 60 anos, atendidas em quatro unidades de Estratégia de Saúde da Família. Resultados: participaram 41 mulheres, com idade média de 52,3 anos. Prevaleceram brancas, casadas, com filhos, ensino médio completo, sem plano de saúde, escore de depressão leve, índice menopausal leve, ansiedade normal e uso regular de medicamentos. Conclusão: as mulheres climatéricas necessitam de um espaço para verbalizar seus sentimentos e dúvidas em relação ao climatério e ter acesso à escuta atenta por profissionais capacitados, visando o atendimento integral de saúde com resolutividade.

Introduction: climacteric can be influenced by simultaneous psychosocial changes, which associated with hormonal changes, such as hypoestrogenism and decreased progesterone levels, can intensify symptoms and health problems in women. Objective: to identify the health profile of climacteric women in primary care regarding their obstetrics, gynecological, social, and demographic aspects, and their lifestyle as well, through a questionnaire and the application of the Hamilton Anxiety Scale, Depression Scale, and the Blatt-Kupperman Menopausal Index. Materials and Methods: it is an exploratory-descriptive cross-sectional study presented as an experience report, developed by medical and nursing students. The participants had 45 to 60 years-old and visited four units of the Family Health Strategy. Results: 41 women participated in the research, with an average age of 52.3-year-old. The majority were white, married, high school graduates, with no health insurance; had children, normal anxiety levels, mild depression, mild menopausal index, and reported regular use of medication. Conclusion: the climacteric women need some time to verbalize their feelings and doubts about the climacteric to trained professionals, aiming for a resolutive and comprehensive health care.

PyJAMAS: open-source, multimodal segmentation and analysis of microscopy images.

SUMMARY: Our increasing ability to resolve fine details using light microscopy is matched by an increasing need to quantify images in order to detect and measure phenotypes. Despite their central role in cell biology, many image analysis tools require a financial investment, are released as proprietary software, or are implemented in languages not friendly for beginners, and thus are used as black boxes. To overcome these limitations, we have developed PyJAMAS, an open-source tool for image processing and analysis written in Python. PyJAMAS provides a variety of segmentation tools, including watershed and machine learning-based methods; takes advantage of Jupyter notebooks for the display and reproducibility of data analyses; and can be used through a cross-platform graphical user interface or as part of Python scripts via a comprehensive application programming interface. AVAILABILITY AND IMPLEMENTATION: PyJAMAS is open-source and available at https://bitbucket.org/rfg_lab/pyjamas. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Management of hereditary angioedema type I and homozygous MTHFR mutation during pregnancy.

Hereditary angioedema (HAE) is an autosomal dominant disease, characterized by edema attacks resulting from quantitative and/or functional deficiency of the C1 inhibitor (C1-INH), which acts in controlling the complement, coagulation, fibrinolysis, and contact systems. The exacerbation of these systems results in decreased circulating levels of kallikrein and conversion of bradykinin. In addition, thrombophilia is related to the deficiency of methylenetetrahydrofolate reductase (MTHFR) enzyme, causing an increase in homocysteine, accumulation of atheromatous plaques, and arterial and venous thrombosis. The association of these conditions in related systems brings greater clinical risks to the patient. We report a female patient, aged 23 years, with HAE and homozygous MTHFR mutation, G2A1, carrier of HAE with crises since early childhood. The first pregnancy terminated with abortion due to gestational sac detachment. In the second pregnancy, at 5.1 weeks, she had bleeding and partial detachment of gestational sac. Thrombophilia tests confirmed homozygosity for the MTHFR enzyme. At the beginning of gestation, she had attacks of angioedema treated with fresh plasma, and at one occasion, she received treatment with a plasma-derived C1-INH esterase. During breastfeeding, she received prophylaxis with plasma-derived C1-INHdp. The course of HAE during pregnancy worsened. There are studies that discuss the occurrence of abortion due to attacks of angioedema. The patient's disease was associated with a homozygous MTHFR mutation, which probably caused the miscarriage. The control of both clinical situations is important for the success of pregnancy, so a combined action plan between obstetricians and HAE treatment specialists is essential.

Management of hereditary angioedema type I and homozygous MTHFR mutation during pregnancy

Hereditary angioedema (HAE) is an autosomal dominant disease, characterized by edema attacks resulting from quantitative and/or functional deficiency of the C1 inhibitor (C1-INH), which acts in controlling the complement, coagulation, fibrinolysis, and contact systems. The exacerbation of these systems results in decreased circulating levels of kallikrein and conversion of bradykinin. In addition, thrombophilia is related to the deficiency of methylenetetrahydrofolate reductase (MTHFR) enzyme, causing an increase in homocysteine, accumulation of atheromatous plaques, and arterial and venous thrombosis. The association of these conditions in related systems brings greater clinical risks to the patient. We report a female patient, aged 23 years, with HAE and homozygous MTHFR mutation, G2A1, carrier of HAE with crises since early childhood. The first pregnancy terminated with abortion due to gestational sac detachment. In the second pregnancy, at 5.1 weeks, she had bleeding and partial detachment of gestational sac. Thrombophilia tests confirmed homozygosity for the MTHFR enzyme. At the beginning of gestation, she had attacks of angioedema treated with fresh plasma, and at one occasion, she received treatment with a plasma-derived C1-INH esterase. During breastfeeding, she received prophylaxis with plasma-derived C1-INHdp. The course of HAE during pregnancy worsened. There are studies that discuss the occurrence of abortion due to attacks of angioedema. The patient’s disease was associated with a homozygous MTHFR mutation, which probably caused the miscarriage. The control of both clinical situations is important for the success of pregnancy, so a combined action plan between obstetricians and HAE treatment specialists is essential (AU)

Contribution of genetic polymorphisms of interleukins IL1B-511 C/T, IL1RN VNTR, IL6-174 G/C, and IL8-251 A/T in gastric lesions: gender and Helicobacter pylori genes matter.

Stomach pathologies develop in a complex interaction between the host's genetic background and H. pylori virulent genes. Thus, our study aimed to compare active chronic gastritis (ACG), and intestinal metaplasia (IM) with inactive chronic gastritis (ICG), according to interleukin polymorphisms of IL6-174 G/C, IL8-251 A/T, IL1ß-511 C/T, and IL1RN VNTR taking into account patient gender and H. pylori genotypes. Interleukin polymorphisms were determined by RFLP-PCR and H. pylori genotype by PCR. IL6-174 GC and IL8-251 T allele showed a protective effect in women against ACG development and, conversely, IL8-251 polymorphism showed a risk for men. More virulent H. pylori strains were associated with the IL8-251 T allele and IL1ß-511 T allele in the AGC, and the vacA m1 allele and cagE gene from H. pylori was associated with the IM. Analysis of the progression of gastric lesions must take into account host variability genetic associated with genes H. pylori due to the relation between the virulent H. pylori genes and more severe gastric lesions, besides the relevance to the gender to IL6-174 and IL8-251 polymorphisms.

Molecular and immunohistochemical analyses of uveal melanoma patient cohort.

Uveal melanoma is a rare form of melanoma and the most frequent primary eye malignancy in adults. The major molecular alterations underlying uveal melanoma pathogenesis affect mainly the GNAQ, GNA11, SF3B1, and BAP1 genes. In this study, we somatically genotyped 31 Brazilian uveal melanomas for BRAF, GNA11, GNAQ, SF3B1, and BAP1 gene mutations and assessed BRCA2 and p53 protein expression. GNAQ and GNA11 mutations were detected in 60%, and SF3B1 mutation rate was 30%. p53 Immunostaining was markedly positive in 5/31, and 3/31 samples showed negative BRCA2 expression. This study supports the importance of these key genes in uveal melanoma tumorigenesis; p53 and BRCA pathways seem to play a role in a subset of patients, possibly heralding unfavorable prognosis.

Cytoskeletal tension regulates mesodermal spatial organization and subsequent vascular fate.

Morphogenesis during human development relies on the interplay between physiochemical cues that are mediated in part by cellular density and cytoskeletal tension. Here, we interrogated these factors on vascular lineage specification during human-induced pluripotent stem-cell (hiPSC) fate decision. We found that independent of chemical cues, spatially presented physical cues induce the self-organization of Brachyury-positive mesodermal cells, in a RhoA/Rho-associated kinase (ROCK)-dependent manner. Using unbiased support vector machine (SVM) learning, we found that density alone is sufficient to predict mesodermal fate. Furthermore, the long-withstanding presentation of spatial confinement during hiPSC differentiation led to an organized vascular tissue, reminiscent of native blood vessels, a process dependent on cell density as found by SVM analysis. Collectively, these results show how tension and density relate to vascular identity mirroring early morphogenesis. We propose that such a system can be applied to study other aspects of the stem-cell niche and its role in embryonic patterning.

A newly validated high-performance liquid chromatography method with diode array ultraviolet detection for analysis of the antimalarial drug primaquine in the blood plasma.

INTRODUCTION:: Primaquine (PQ) diphosphate is an 8-aminoquinoline antimalarial drug with unique therapeutic properties. It is the only drug that prevents relapses of Plasmodium vivax or Plasmodium ovale infections. In this study, a fast, sensitive, cost-effective, and robust method for the extraction and high-performance liquid chromatography with diode array ultraviolet detection (HPLC-DAD-UV ) analysis of PQ in the blood plasma was developed and validated. METHODS:: After plasma protein precipitation, PQ was obtained by liquid-liquid extraction and analyzed by HPLC-DAD-UV with a modified-silica cyanopropyl column (250mm × 4.6mm i.d. × 5µm) as the stationary phase and a mixture of acetonitrile and 10mM ammonium acetate buffer (pH = 3.80) (45:55) as the mobile phase. The flow rate was 1.0mL·min-1, the oven temperature was 50OC, and absorbance was measured at 264nm. The method was validated for linearity, intra-day and inter-day precision, accuracy, recovery, and robustness. The detection (LOD) and quantification (LOQ) limits were 1.0 and 3.5ng·mL-1, respectively. The method was used to analyze the plasma of female DBA-2 mice treated with 20mg.kg-1 (oral) PQ diphosphate. RESULTS:: By combining a simple, low-cost extraction procedure with a sensitive, precise, accurate, and robust method, it was possible to analyze PQ in small volumes of plasma. The new method presents lower LOD and LOQ limits and requires a shorter analysis time and smaller plasma volumes than those of previously reported HPLC methods with DAD-UV detection. CONCLUSIONS:: The new validated method is suitable for kinetic studies of PQ in small rodents, including mouse models for the study of malaria.

DNA methylation polymerase chain reaction (PCR) array of apoptosis-related genes in pleomorphic adenomas of the salivary glands.

OBJECTIVE: The aim of this study was to evaluate the DNA methylation profile in 22 apoptosis-related genes in pleomorphic adenomas (PAs) of the salivary glands, in comparison with normal salivary glands (NSGs), and to address the differences in methylation patterns between smaller and larger tumors. Additionally, we investigated if the hypermethylation of differentially methylated genes between NSGs and PAs impacted the messenger RNA (mRNA) transcription. DESIGN: Twenty-three fresh PA samples and 12 NSG samples were included. The PA samples were divided into 2 groups: PAs with clinical size larger than 2 cm (n = 12) and PAs with clinical size 2 cm or smaller (n = 11). DNA methylation at the promoter region of a panel of 22 genes involved in apoptosis was profiled by using a human apoptosis DNA methylation polymerase chain reaction array, and the transcriptional levels of genes showing differential methylation profiles between PAs and NSGs were assessed. RESULTS: TNFRSF25 and BCL2 L11 were highly methylated in PAs, in comparison with NSGs, irrespective of tumor size. However, no difference could be observed in the mRNA transcription between PAs and NSGs. CONCLUSIONS: Hypermethylation of the proapoptotic genes BCL2 L11 and TNFRSF25 is observed in PA. However, this phenomenon did not impact mRNA transcription.

A newly validated high-performance liquid chromatography method with diode array ultraviolet detection for analysis of the antimalarial drug primaquine in the blood plasma

Abstract INTRODUCTION: Primaquine (PQ) diphosphate is an 8-aminoquinoline antimalarial drug with unique therapeutic properties. It is the only drug that prevents relapses of Plasmodium vivax or Plasmodium ovale infections. In this study, a fast, sensitive, cost-effective, and robust method for the extraction and high-performance liquid chromatography with diode array ultraviolet detection (HPLC-DAD-UV ) analysis of PQ in the blood plasma was developed and validated. METHODS: After plasma protein precipitation, PQ was obtained by liquid-liquid extraction and analyzed by HPLC-DAD-UV with a modified-silica cyanopropyl column (250mm × 4.6mm i.d. × 5μm) as the stationary phase and a mixture of acetonitrile and 10mM ammonium acetate buffer (pH = 3.80) (45:55) as the mobile phase. The flow rate was 1.0mL·min-1, the oven temperature was 50OC, and absorbance was measured at 264nm. The method was validated for linearity, intra-day and inter-day precision, accuracy, recovery, and robustness. The detection (LOD) and quantification (LOQ) limits were 1.0 and 3.5ng·mL-1, respectively. The method was used to analyze the plasma of female DBA-2 mice treated with 20mg.kg-1 (oral) PQ diphosphate. RESULTS: By combining a simple, low-cost extraction procedure with a sensitive, precise, accurate, and robust method, it was possible to analyze PQ in small volumes of plasma. The new method presents lower LOD and LOQ limits and requires a shorter analysis time and smaller plasma volumes than those of previously reported HPLC methods with DAD-UV detection. CONCLUSIONS: The new validated method is suitable for kinetic studies of PQ in small rodents, including mouse models for the study of malaria.

Compliant substratum guides endothelial commitment from human pluripotent stem cells.

The role of mechanical regulation in driving human induced pluripotent stem cell (hiPSC) differentiation has been minimally explored. Although endothelial cell (EC) fate from hiPSCs has been demonstrated using small molecules to drive mesoderm induction, the effects of substrate stiffness with regard to EC differentiation efficiency have yet to be elucidated. We hypothesized that substrate compliance can modulate mesoderm differentiation kinetics from hiPSCs and affect downstream EC commitment. To this end, we used polydimethylsiloxane (PDMS)-a transparent, biocompatible elastomeric material-as a substrate to study EC commitment of hiPSCs using a stepwise differentiation scheme. Using physiologically stiff (1.7 MPa) and soft (3 kPa) PDMS substrates, compared to polystyrene plates (3 GPa), we demonstrate that mechanical priming during mesoderm induction activates the Yes-associated protein and drives Wnt/ß-catenin signaling. When mesoderm differentiation was induced on compliant PDMS substrates in both serum and serum-free E6 medium, mesodermal genetic signatures (T, KDR, MESP-1, GATA-2, and SNAIL-1) were enhanced. Furthermore, examination of EC fate following stiffness priming revealed that compliant substrates robustly improve EC commitment through VECad, CD31, vWF, and eNOS marker expression. Overall, we show that substrate compliance guides EC fate by enhancing mesoderm induction through Wnt activation without the addition of small molecules. These findings are the first to show that the mechanical context of the differentiation niche can be as potent as chemical cues in driving EC identity from hiPSCs.

Main Reasons for Registration Application Refusal of Generic and Similar Pharmaceutical Drug Products by the Brazilian Health Regulatory Agency (ANVISA).

Objective. The marketing authorization of generic and similar pharmaceutical drug products involves the analysis of proposing company's administrative aspects as well as drug product technical description and scientific evaluations. This study evaluated the main reasons for registration refusal of generic and similar pharmaceutical drug products in Brazil. The aim is to help future applicants to better organize the proposal. Methods. A retrospective search of drug products registration processes was performed on the Brazilian Government Official Gazette from January 1, 2015, and December 31, 2015. Results. Drug product quality control, drug product stability study, deadline accomplishment, API quality control made by drug manufacturer, active pharmaceutical ingredient (API), and production report were the main reasons for marketing authorization application refusal of generic and similar pharmaceutical drug products in 2015. Conclusion. Disclosure of the reasons behind failed applications is a step forward on regulatory transparency. Sharing of experiences is essential to international regulatory authorities and organizations to improve legislation requirements for the marketing authorization of generic and similar pharmaceutical drug products.

Fatores associados à venda da Agricultura Familiar para o Programa Nacional de Alimentação Escolar nos municípios do estado de São Paulo / Associated factors with the sale of the Family Farming for the Brazilian School Feeding Program in the cities of São Paulo State

Introdução: A agricultura familiar (AF) e o PNAE após um processo de evolução política se vincularam em 2009 por meio da Lei n°11.947. O agricultor familiar a partir desse momento passou a dispor de um importante mercado institucional. Entretanto, esse ator apresenta dificuldades em atender à demanda do PNAE. Objetivo: Analisar os fatores associados à venda da AF para o PNAE nos municípios do estado de São Paulo em 2012. Metodologia: Trata-se de estudo transversal, envolvendo métodos de análise quantitativa. Os dados analisados foram coletados da pesquisa O encontro entre o Programa Nacional de Alimentação Escolar (PNAE) e a agricultura familiar: avaliação do processo de implementação e dos possíveis efeitos das compras locais, segundo a Lei n°11.947/2009, a qual avaliou 38 municípios do Estado de São Paulo. Foram coletados dados de 110 agricultores que venderam para o PNAE. Os eixos de variáveis estudadas foram: características socioeconômicas dos municípios, características do agricultor, apoio institucional e característica da venda. O percentual da compra da AF para o PNAE = 30 por cento foi a variável desfecho do estudo. A descrição e caracterização foram calculadas por meio de medidas de tendência central e dispersão e frequências absolutas e relativas. O teste de McNemar foi utilizado para verificar diferenças entre a comercialização antes e depois da Lei n° 11.947/09. Foi feita uma regressão simples bivariada com o intuito de verificar o incremento no percentual da compra da AF para o PNAE entre 2011 e 2015. E foi utilizada a análise de regressão logística múltipla para analisar os fatores associados à venda da AF para o PNAE. Resultados: Os municípios estudados apresentaram IDHM alto (87 por cento ); os alimentos produzidos pelos agricultores eram em sua quase totalidade in natura; os agricultores depois da Lei n°11.947/2009 aumentaram suas vendas para os mercados institucionais; a média no percentual da compra da agricultura familiar nos 16 municípios estudados subiu de 25,3 por cento para 36,3 por cento entre os anos de 2011 a 2015. Os municípios com PIB mais alto e que já apresentavam uma experiência com o Programa de Aquisição de Alimentos (PAA) se associaram significativamente com a venda da AF para o PNAE exigida pela Lei n°11.947/2009. No plano do agricultor, àqueles que obtiveram a Declaração de Aptidão ao PRONAF (DAP) depois da Lei n°11.947/2009 se associaram significativamente com o desfecho, OR 3,20 (1,19 8,63); os agricultores que vendiam para o PAA antes da Lei se associaram ao desfecho com OR 0,09 (0,03 0,33). O apoio da nutricionista se associou com o desfecho com OR 3,76 (1,18 12,03). Os agricultores não individuais (associados e cooperados) não se associaram com o desfecho percentual da compra = 30 por cento . Conclusão: Os municípios com PIB mais alto, em que já apresentavam uma experiência com o PAA, os agricultores que obtiveram a DAP após a Lei, o suporte técnico da nutricionista são fatores importantes associados à venda da AF para o PNAE

Introduction: The Family Farming (FF) and The Brazilian School Feeding Program (PNAE) after a process of political evolution were associated in 2009 by Brazilian act 11.947/2009. Since that time, the family farmer has an important institutional market available. However, this actor has difficulties to achieve the market demand. Objective: To analyze the factors associated to the sale of FF to the PNAE in the cities of the state of São Paulo in 2012. Methodology: The study is transverse, using quantitative analysis methods. The analyzed data were collected from the study \"The meeting of the National School Feeding Program and Family Farming: evaluation of the implementation process and the possible effects of local purchases, according to the act 11.947/2009, which evaluated 38 cities in the São Paulo state. Data were collected from 110 family farmers who sold to the PNAE. The main variables studied were: cities socioeconomic characteristics, farmer characteristics, institutional support and sale characteristics. The FF purchase percentage for the PNAE = 30 per cent was the outcome variable of the study. The description and characterization were calculated through measures of central tendency and dispersion and absolute and relative frequencies. McNemar´s test was used to verify differences between sales before and after the act 11.947/09. A simple bivariate linear regression was performed in order to verify the increase in the percentage of the FF purchases for the PNAE between 2011 and 2015. The multiple logistic regression analysis was used to analyze the factors associated to the FF sale to PNAE. Results: The studied cities had high Human Development Index (HDI) (87 per cent ); the produced food by family farmers was almost entirely in natura; the family farmers after the act 11.947/09 increased their sales to the institutional markets; the average percentage of FF purchases in the 16 studies cities increased from 25.3 per cent to 36.3 per cent between 2011 and 2015. The cities with the highest Gross Domestic Product (GDP) and previous experience with the Family Agriculture Food Acquisition Program (PAA) were significantly associated to the FF sale for the BSFP required by Brazilian Law. At the farmer\'s level, those who obtained Ability Declaration for National Program for the Strengthening of Family Farming (DAP) after the Brazilian act were significantly associated with the outcome, OR 3,20 (1,19 8,63); the family farmers who sold to the PAA before the act were associated with the outcome OR 0,09 (0,03 0,33). The nutritionist\'s support was associated with the outcome OR 3,76 (1,18 12,03). Non-individual farmers were not significantly associated with the outcome. Conclusion: Cities with: high GDP, previous experience with the PAA, family farmers with DAP experience, technical nutritionist support are important factors associated to the sale of FF for the PNAE