Testes bioquímicos para identificação de leveduras na medicina veterinária - revisão de literatura / Biochemical tests to yeast identification in veterinary medicine - literature review / Pruebas bioquímicas para identificación de levaduras en la medicina veterinaria - revisión de literatura

As leveduras são fungos de importância à medicina veterinária por causarem doenças infecciosas em diferentes hospedeiros animais. A presente revisão de literatura teve como objetivo relatar os principais testes bioquímicos capazes de auxiliar na identificação de fungos leveduriformes de interesse veterinário e zoonótico. Para o levantamento bibliográfico, foram consideradas 48 publicações científicas selecionadas na área e indexadas nas principais bases de dados, entre os anos de 1988 e 2020. Como resultados, observou-se que oito provas são as mais empregadas na rotina micológica. Devido à baixa variabilidade morfológica das espécies leveduriformes, testes bioquímicos complementares são fundamentais na rotina laboratorial. A análise do perfil bioquímico de leveduras contribui na determinação taxonômica dos fungos a partir de reações químicas, visto que o metabolismo varia de acordo com a espécie, resultando em metabólitos distintos, os quais podem ser avaliados por diferentes provas. Conclui-se que a identificação fenotípica das leveduras é imprescindível no diagnóstico, prognóstico, tratamento e controle de doenças fúngicas e contribui para a manutenção da saúde animal.(AU)

Yeasts are fungi of importance to veterinary medicine because they cause infectious diseases in different animal hosts. This literature review aimed to report the main biochemical tests capable of assisting in the identification of yeast-like fungi of veterinary and zoonotic interest. For the bibliographical survey, 48 selected scientific publications in the area and indexed in the main databases, between the years 1988 and 2020, were considered. As a result, it was observed that eight tests are the most used in the mycological routine. Due to the low morphological variability of yeast species, complementary biochemical tests are fundamental in the laboratory routine. The analysis of the biochemical profile of yeast contributes to the taxonomic determination of fungi based on chemical reactions, since the metabolism varies according to the species, resulting in different metabolites, which can be evaluated by different tests. It is concluded that the phenotypic identification of yeasts is essential in the diagnosis, prognosis, treatment and control of fungal diseases and contributes to the maintenance of animal health.(AU)

Las levaduras son hongos de importancia para la medicina veterinaria porque causan enfermedades infecciosas en diferentes animales huéspedes. Esta revisión de la literatura tuvo como objetivo informar las principales pruebas bioquímicas capaces de ayudar en la identificación de hongos tipo levadura de interés veterinario y zoonótico. Para el levantamiento bibliográfico se consideraron 48 publicaciones científicas seleccionadas en el área e indexadas en las principales bases de datos, entre los años 1988 y 2020. Como resultado se observó que ocho pruebas son las más utilizadas en la rutina micológica. Debido a la baja variabilidad morfológica de las especies de levaduras, las pruebas bioquímicas complementarias son fundamentales en la rutina del laboratorio. El análisis del perfil bioquímico de la levadura contribuye a la determinación taxonómica de los hongos en base a reacciones químicas, ya que el metabolismo varía según la especie, dando como resultado diferentes metabolitos, los cuales pueden ser evaluados mediante diferentes pruebas. Se concluye que la identificación fenotípica de levaduras es fundamental en el diagnóstico, pronóstico, tratamiento y control de enfermedades fúngicas y contribuye al mantenimiento de la salud animal.(AU)

Use of cerebrospinal fluid CXCL13 concentration for diagnosis and monitoring of neurosyphilis: a three-case report

Introduction: Previous retrospective studies have demonstrated that the concentration of chemokine ligand CXCL13 in cerebrospinal fluid (CSF-CXCL13) is a promising biomarker in the diagnosis of neurosyphilis and, additionally, in the monitoring of therapeutic efficacy. Objective: To describe three cases of patients with neurosyphilis (NS) treated at Hospital Universitário Gaffrée e Guinle, in Rio de Janeiro, Brazil, with suspected active syphilis with neurological symptoms. Case report: Three patients from Rio de Janeiro, Brazil, were investigated for symptomatic NS. The concentration of CSF-CXCL13 was prospectively performed by enzyme-linked immunosorbent assay (ELISA) in all participants at baseline and in follow-up visits at 3 months after therapy. CSF-CXCL13 concentrations were significantly higher in all three patients with established NS. The CSF-CXCL13 concentrations decreased after 3 months of therapy compared to baseline in all cases reported. The added high concentration of CSF-CXCL13 plus CSF-TPHA reactivity above 1:40 titer agreed with the diagnosis of NS in 100% of the cases. Conclusion: In this case series, we present three cases of NS diagnosed using CXCL13 in CSF as a complementary test. These case series suggest that the clinical use of CSF-CXCL13 is useful as a supplementary biomarker for NS and for monitoring the effectiveness of NS therapy, especially in patients with nonreactive CSF-VDRL, excluding other neurologic diseases

Periprosthetic gout flare after total knee arthroplasty: A misdiagnostic case report / 北京大学学报(医学版)

Periprosthetic gout flare is a rare arthritic condition after total knee arthroplasty, but the symptoms of gout may have often been mistaken as acute periprosthetic infection given their similarity. Misdiagnosis as periprosthetic infection can lead to unnecessary surgery, long-term dependence on anti-biotics, and even malfunction of the involved knee joint. Here, we report a case study of a patient with immunodeficiency condition of long-term oral glucocorticoid and diabetes mellitus, who had undergone a knee replacement 8 weeks before. The initial symptoms of fever and joint pain together with the dysfunction of her right knee with elevated inflammatory markers, such as increased serum leukocytes, erythrocyte sedimentation rate, C-reactive protein, and synovial cell counts led to a diagnosis of acute periprosthetic infection. Arthrocentesis and bacterial culture were performed preoperatively. According to the current Musculoskeletal Infection Society (MSIS) criteria for diagnosis of periprosthetic infection, the case was classified as periprosthetic infection and a prosthesis retained debridement surgery was performed. However we got negative culture results in all the pre-operative and intro-operative samples. The symptoms as well as the laboratory inflammatory markers improved shortly after the debridement surgery until the 11th day when all the similar systemic and local symptoms recurred. With a remedial crystal analysis of synovial fluid from the patient, gouty flare was found to be the cause of acute arthritis finally. Accor-dingly, after anti-gout medications were administrated, the symptoms associated with acute arthritis gra- dually subsided, and there was no recurrence during a 24-month follow-up. This article described the cli-nical manifestation, diagnosis and differential diagnosis, treatment of a case of periprosthetic gout. Although relatively rare, gout should be considered as a differential diagnosis in suspected periprosthetic infection. Current criteria for periprosthetic infection can not exclude the diagnosis of periprosthetic gout flare, it is therefore imperative that the analysis of joint aspirate for crystals be conducted to determine the correct course of treatment, or unnecessary surgical procedure may be performed in periprosthetic gout case.

Value of glycosylated hemoglobin A1c and apolipoprotein A-1 ratio on predicting outcome of patients with acute coronary syndrome / 中华心血管病杂志

Objective: To investigate the predictive value of glycosylated hemoglobin A1c/apolipoprotein A-1 (HbA1c/ApoA-1) ratio for major adverse cardiovascular events (MACEs) in patients with acute coronary syndrome (ACS). Methods: The present study is a retrospective cohort study. ACS patients who were hospitalized and underwent coronary angiography at Beijing Hospital from March 2017 to March 2019 were enrolled. Baseline information such as sex, age, previous history, Gensini score, HbA1c and ApoA-1 were analyzed. Patients were divided into two groups according to presence or absence of MACEs and the difference on HbA1c/ApoA-1 ratio was compared between the two groups. According to the tertiles of HbA1c/ApoA-1 levels, patients were divided into high (5.87-16.12), medium (4.50-5.83) and low (2.11-4.48) HbA1c/ApoA-1 groups. Cox proportional risk model was used to evaluate the differences in MACEs and all-cause mortality among the three groups. Kaplan-Meier survival analysis was used to compare the differences of MACEs between the various HbA1c/ApoA-1 groups. Results: A total of 366 ACS patients were included in this study. The mean age of the patients was (65.9±10.3) years. There were 59 MACEs and 10 all-cause deaths during the mean of (22.3±4.4) months follow-up. After adjusting for age, systolic blood pressure, history of diabetes and Gensini score, the incidence of MACEs was 2.45 times higher in the high HbA1c/ApoA-1 group than in the low HbA1c/ApoA-1 group (95%CI 1.16-5.18, P=0.019). There was no significant difference in all-cause mortality between the high and low HbA1c/ApoA-1 groups (P=1.000). Kaplan-Meier survival analysis showed that patients in the high HbA1c/ApoA-1 group had the highest risk of MACEs, while patients in the low HbA1c/ApoA-1 group had the lowest risk of MACEs (P<0.01). Spearman rank correlation analysis showed that HbA1/ApoA-1 ratio was positively correlated with Gensini score in ACS patients (r=0.274, P<0.01). Conclusion: High HbA1c/ApoA-1 ratio was an independent risk factor for MACEs in ACS patients. Patients with high HbA1c/ApoA-1 ratio had more severe coronary artery disease lesions. HbA1c/ApoA-1 ratio may be used as a potential risk stratification biomarker for ACS patients, it might be useful for the early identification of high-risk population and for predicting the incidence of MACEs among ACS patients.

Clinical relevance of oxidative stress biomarkers in cardiovascular prevention / Relevância clínica de biomarcadores de estresse oxidativo na prevenção cardiovascular

Cardiovascular diseases involve hyperlipidemia, inflammation and oxidative stress. Although this relationship is well established, only biomarkers associated with hyperlipidemia and inflammation are currently in clinical practice for diagnosis and evaluation of patient treatment. Our hypothesis is that oxidative stress biomarkers may be an independent risk factor and may assist in cardiovascular risk stratification and contribute to improving current scores. Thus, the objective of this study was to investigate which are the biomarkers and methodologies were used in clinical studies in humans with different health conditions. With the results obtained in the first part, we selected studies conducted in healthy individuals and in individuals under primary and secondary cardiovascular prevention in order to evaluate the most frequent biomarkers, the results obtained according to the individual's profile and the methodology used, and correlate with different health conditions. We observed that malondialdehyde (MDA) was the most frequent lipid biomarker of oxidative stress applied in the studies, but it presented significant variability in the results and a weak correlation with clinical outcomes. The result of this study demonstrates the importance of carrying out a multicentric study to validate the MDA values in individuals with different health conditions and the standardization of the methodology based on high performance liquid chromatographyy (HPLC)

As doenças cardiovasculares envolvem hiperlipidemia, inflamação e estresse oxidativo. Embora essa relação esteja bem estabelecida, apenas biomarcadores associados à hiperlipidemia e inflamação são atuais na prática clínica para diagnóstico e avaliação do tratamento do paciente. Nossa hipótese é que biomarcadores de estresse oxidativo podem ser um fator de risco independente e podem auxiliar na estratificação de risco cardiovascular e contribuir para melhorar os escores atuais. Assim, o objetivo deste estudo foi investigar primeiramente quais são os biomarcadores e metodologias utilizados nos estudos clínicos em humanos em diferentes condições de saúde. Com os resultados obtidos na primeira etapa, selecionamos os estudos conduzidos em indivíduos saudáveis e em prevenção cardiovascular primária e secundária a fim de avaliar os biomarcadores mais utilizados, os resultados obtidos conforme o perfil do indivíduo e a metodologia utilizada e finalmente correlacionar com as diferentes condições de saúde. Observamos que o malondialdeído (MDA) foi o biomarcador lipídico de estresse oxidativo mais frequente nos estudos, porém apresentou importante variabilidade nos resultados e fraca correlação com desfechos clínicos. O resultado desse estudo demonstra a importância da realização de um estudo multicentrico para validação dos valores de MDA nos diferentes perfis de indivíduos e a padronização metodológica baseada na cromatografia líquida de alta eficiência (HPLC)

Advancements in artificial intelligence and machine learning in revolutionising biomarker discovery

Abstract The article explores the significance of biomarkers in clinical research and the advantages of utilizing artificial intelligence (AI) and machine learning (ML) in the discovery process. Biomarkers provide a more comprehensive understanding of disease progression and response to therapy compared to traditional indicators. AI and ML offer a new approach to biomarker discovery, leveraging large amounts of data to identify patterns and optimize existing biomarkers. Additionally, the article touches on the emergence of digital biomarkers, which use technology to assess an individual's physiological and behavioural states, and the importance of properly processing omics and multi-omics data for efficient handling by computer systems. However, the article acknowledges the challenges posed by AI/ML in the identification of biomarkers, including potential biases in the data and the need for diversity in data representation. To address these challenges, the article suggests the importance of regulation and diversity in the development of AI/ML algorithms.

Comparative Assessment of E-cadherin's Expression between the Metastatic and Non-metastatic Oral Squamous Cell Carcinoma: An Immunohistochemical Study

ABSTRACT Objective: To identify the clinicopathological correlation of E-cadherin expression in metastatic and non-metastatic oral squamous cell carcinoma (OSCC). Material and Methods: A total of 90 paraffin-embedded tissue sections of OSCC were retrieved from the registry. The total selected samples were 45 cases each from the primary lesions of metastatic and non-metastatic OSCC. One section was subjected to routine Hematoxylin and eosin stain and another to immunohistochemical analysis for E-cadherin expression. Results: A non-significant (p˃0.05) increased expression is seen in the non-metastatic group compared to the metastatic group, with predominantly membrane as the staining site in either group. However, the expression of E-cadherin did not reveal any statistically significant association with independent variables such as age, gender, and adverse habits of the patients (p>0.05). On the other hand, with respect to the histological differentiation of OSCC, a significant association (p<0.001) was observed with the well-differentiated type of metastatic OSCC. Conclusion: E-cadherin was useful to some extent in predicting regional metastasis. However, further studies using a panel of biomarkers with increased sample size may help us understand the process involved in metastasis.

Cell phenotypes as activity biomarkers in patients with Systemic Lupus Erythematosus

Abstract The pathogenesis of systemic lupus erythematosus (SLE) is complex. Few studies in Brazilian population have addressed cell phenotypes associated with immunological responses and their associations with SLE activity. The aim of this study is to investigate cell phenotypes associated to SLE diagnosis, treatment and activity. Twenty-eight SLE female patients (17 inactive, 11 active) and 10 healthy women were included in this study. Markers of natural killer (Nk), T and B cells in peripheral blood were evaluated by flow cytometry. Nkt cells were decreased only in SLE active patients. Activated CD4+, regulatory T FoxP3+ and B cells were decreased in both active and inactive SLE patients, compared to control group. The data corroborate the disruption of immune regulatory response in SLE patients and suggest phenotipic changes as possible biomarkers of SLE activity.

Analysis and research on fitness biomarkers in volleyball athletes during competition / Análise e pesquisa sobre biomarcadores de condicionamento físico em atletas de vôlei durante competição / Análisis e investigación de los biomarcadores de aptitud física en atletas de voleibol durante competición

ABSTRACT Introduction: Athletes' physical conditioning is closely related to the type of sport dedicated, training levels, intrinsic conditions of the athletes, type of load, and recovery status after exercise training. There is ample documentation on the physical conditioning of volleyball athletes in normal situations, but the literature is lacking in surveys during competition periods. Objective: To explore the physical conditioning through analysis of body biomarkers in volleyball athletes in competition. Methods: Before and after the summer training, fasting urine and blood samples were collected from five female volleyball players. Biomarkers involving white blood cells, red blood cells, hemoglobin, etc., were detected, recorded, and the data were statistically analyzed. After the general analysis of the sample, a case study of two elite female athletes was conducted. Results: The urea nitrogen value of athlete A decreased from 7.62mmol/L to 4.91mmol/L, and the urea nitrogen value of athlete B decreased from 6.91mmol/L to 6.57mmol/L; the urea nitrogen values of both athletes showed a downward trend. Athlete A's physical function status decreased significantly after summer training, while Athlete B's physical function status improved slightly. Conclusion: During the summer training period, it is necessary to perform continuous index measurements on the athletes to increase the timeliness of the data, which is conducive to in-depth analysis of the athletes' fitness status. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: O condicionamento físico dos atletas está intimamente relacionado ao tipo de esporte dedicado, níveis de treinamento, condições intrínsecas das atletas, tipo de carga e status de recuperação após o treinamento dos exercícios. Há vasta documentação sobre o condicionamento físico dos atletas de vôlei em situações normais, porém a literatura deixa a desejar nos levantamentos durante o período de competições. Objetivo: Explorar o condicionamento físico através de análise de biomarcadores corporais nos atletas de vôlei em competição. Métodos: Antes e depois do período de treinamento de verão, foram coletadas amostras de urina e sangue, em jejum, de cinco jogadoras de vôlei feminino. Biomarcadores envolvendo glóbulos brancos, glóbulos vermelhos, hemoglobina, etc. foram detectados, registrados, e os dados foram analisados estatisticamente. Após a análise geral da amostra, foi realizado um estudo de caso de duas atletas de elite. Resultados: O valor de nitrogênio de ureia da atleta A diminuiu de 7,62mmol/L para 4,91mmol/L, e o valor do nitrogênio de ureia da atleta B diminuiu de 6,91mmol/L para 6,57mmol/L, os valores de nitrogênio da ureia de ambas as atletas apresentaram tendência de queda. O estado de função física da atleta A diminuiu significativamente após os treinos de verão, enquanto o estado de função física da atleta B melhorou ligeiramente. Conclusão: Durante o período de treinamento de verão, é necessário realizar uma medição contínua do índice sobre os atletas para aumentar a pontualidade dos dados, o que é propício à análise aprofundada do estado de condicionamento físico das atletas. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: El acondicionamiento físico de los deportistas está íntimamente relacionado con el tipo de deporte al que se dedican, los niveles de entrenamiento, las condiciones intrínsecas de los deportistas, el tipo de carga y el estado de recuperación tras los ejercicios de entrenamiento. Existe una amplia documentación sobre el acondicionamiento físico de los deportistas de voleibol en situaciones normales, pero la literatura deja mucho que desear en los estudios durante el periodo de competiciones. Objetivo: Explorar la condición física mediante el análisis de biomarcadores corporales en atletas de voleibol en competición. Métodos: Antes y después del periodo de entrenamiento de verano, se recogieron muestras de orina y sangre en ayunas de cinco jugadoras de voleibol. Se detectaron y registraron los biomarcadores relativos a los glóbulos blancos, los glóbulos rojos, la hemoglobina, etc., y los datos se analizaron estadísticamente. Tras el análisis general de la muestra, se realizó un estudio de casos de dos atletas de élite. Resultados: El valor de nitrógeno ureico del atleta A disminuyó de 7.62mmol/L a 4.91mmol/L, y el valor de nitrógeno ureico del atleta B disminuyó de 6.91mmol/L a 6.57mmol/L, los valores de nitrógeno ureico de ambos atletas mostraron una tendencia a la baja. El estado de la función física del atleta A disminuyó significativamente después del entrenamiento de verano, mientras que el estado de la función física del atleta B mejoró ligeramente. Conclusión: Durante el período de entrenamiento de verano, es necesario realizar mediciones continuas de los índices en los atletas para aumentar la puntualidad de los datos, lo que favorece el análisis en profundidad del estado de forma física de los atletas.Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Relación de IMC con marcadores de inflamación y hemoglobina glicosilada en niños y jóvenes con diabetes mellitus 1 / Relationship of BMI with markers of inflammation and glycosylated hemoglobin in children and young adults with diabetes mellitus 1

La diabetes mellitus tipo 1 (DM1) es una enfermedad autoinmune que genera dependencia exógena de insulina de forma permanente, presenta inflamación subclínica crónica lo que conlleva a una elevación de marcadores de inflamación como factor de necrosis tumoral alfa (TNF-α), proteína C reactiva (PCR) e interleuquina 6 (IL-6). OBJETIVO: determinar la relación entre el IMC sobre los marcadores de inflamación y el control metabólico en niños y jóvenes con DM1 entre 5 a 15 años de edad. METODOLOGÍA: Se realizó un estudio clínico, observacional, exploratorio. A partir de La recolección de datos de fichas clínicas y muestras de sangre en el Instituto de Investigaciones Materno Infantil (IDIMI) del Hospital San Borja Arriarán de la Universidad de Chile. Clasificación del estado nutricional utilizando datos registrados en ficha clínica. Marcadores de inflamación por medio de ELISA, hemoglobina glicosilada mediante métodos estándares. El análisis estadístico incluyó correlaciones mediante test de Spearman y diferencia de medias mediante test de Kruskal-Wallis seguido de post hoc Dunns. RESULTADOS: Un 30% de los pacientes con DM1 presentaron malnutrición por exceso. Al analizar la relación entre los niveles de marcadores inflamatorios y Hb glicosilada se observó la existencia de asociacion positiva entre usPCR y HbA1c (r= 0,30; p=0,0352) y entre IL-6 y HbA1c (r= - 0,038; p=0,0352). CONCLUSIONES: este estudio describe una posible asociación entre parámetros clásicos de inflamación con la hemoglobina glicosilada en las categorias de sobrepeso y obesidad en pacientes con DM1.

Type 1 diabetes mellitus (T1D) is an autoimmune disease that generates permanent exogenous insulin dependence, accompanied by chronic subclinical inflammation that leads to an elevation of inflammation markers such as tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP) and interleukin-6 (IL-6). OBJECTIVE: To determine the relationship between BMI on markers of inflammation and metabolic control in children and young people with T1D between 5 and 15 years of age. METHODOLOGY: A clinical, observational and exploratory study was carried out, based on the collection of data from clinical records and blood samples of children and adolescents with DM1 at the Instituto de Investigaciones Materno Infantil (IDIMI) of the Hospital San Borja Arriarán of the Universidad de Chile. Nutritional status, levels of inflammation markers and glycosylated hemoglobin were determined by standardized methods. Statistical analysis included correlations by Spearman test and mean difference by Kruskal-Wallis test followed by post hoc Dunns test. RESULTS: A total of 56 patients with T1D were analyzed, 30% of whom presented excess malnutrition. Those children or adolescents with obesity presented significantly higher usPCR levels compared to underweight patients or patients at risk of malnutrition (p=0.039). In addition, HbA1c levels were determined which were negatively associated with usPCR (r= 0.30; p=0.0352) and IL-6 (r= - 0.038; p=0.0352) levels. CONCLUSIONS: This study points out that nutritional status is associated with usPCR levels, in agreement with what is described in the literature and shows a possible association between classical parameters of inflammation with glycosylated hemoglobin in children and adolescents with nutritional diagnosis of overweight or obesity.

Deep learning for analysis of changes in vital capacity and blood markers after swimming matches based on blended learning / Aprendizado profundo para análise das alterações da capacidade vital e marcadores sanguíneos após jogos de natação baseados no aprendizado combinado / Aprendizaje profundo para el análisis de los cambios en la capacidad vital y los marcadores sanguíneos después de los partidos de natación basados en el aprendizaje combinado

ABSTRACT Introduction Nowadays, more people are concerned with physical exercise and swimming competitions, as a major sporting event, have become a focus of attention. Such competitions require special attention to their athletes and the use of computational algorithms assists in this task. Objective To design and validate an algorithm to evaluate changes in vital capacity and blood markers of athletes after swimming matches based on combined learning. Methods The data integration algorithm was used to analyze changes in vital capacity and blood acid after combined learning swimming competition, followed by the construction of an information system model to calculate and process this algorithm. Results Comparative experiments show that the neural network algorithm can reduce the calculation time from the original initial time. In the latest tests carried out in about 10 seconds, this has greatly reduced the total calculation time. Conclusion According to the model requirements of the designed algorithm, practical help has been demonstrated by building a computational model. The algorithm can be optimized and selected according to the calculation model according to the reality of the application. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução Atualmente, mais pessoas preocupam-se com o exercício físico e as competições de natação, como evento esportivo de destaque, tornou-se foco de atenção. Tais competições exigem atenção especial aos seus atletas e o uso de algoritmos computacionais auxiliam nessa tarefa. Objetivo Projetar e validar um algoritmo para avaliação das alterações da capacidade vital e marcadores sanguíneos dos atletas após os jogos de natação baseados no aprendizado combinado. Métodos O algoritmo de integração de dados foi usado para analisar as mudanças de capacidade vital e ácido sanguíneo após competição de natação de aprendizado combinado, seguido à construção de um modelo de sistema de informação para calcular e processar esse algoritmo. Resultados Experiências comparativas mostram que o algoritmo de rede neural pode reduzir o tempo de cálculo a partir do tempo inicial original. Nos últimos testes levados à cabo em cerca de 10 segundos, isto reduziu muito o tempo total de cálculo. Conclusão De acordo com os requisitos do modelo do algoritmo projetado, foi demonstrada a ajuda prática pela construção de um modelo computacional. O algoritmo pode ser otimizado e selecionado de acordo com o modelo de cálculo, segundo a realidade da aplicação. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción Hoy en día, cada vez más personas se preocupan por el ejercicio físico y las competiciones de natación, como evento deportivo destacado, se han convertido en un foco de atención. Estas competiciones requieren una atención especial para sus atletas y el uso de algoritmos computacionales ayuda en esta tarea. Objetivo Diseñar y validar un algoritmo para evaluar los cambios en la capacidad vital y los marcadores sanguíneos de los atletas después de los partidos de natación basado en el aprendizaje combinado. Métodos Se utilizó el algoritmo de integración de datos para analizar los cambios de la capacidad vital y la acidez de la sangre tras la competición de natación de aprendizaje combinado, seguido de la construcción de un modelo de sistema de información para calcular y procesar este algoritmo. Resultados Los experimentos comparativos muestran que el algoritmo de la red neuronal puede reducir el tiempo de cálculo con respecto al tiempo inicial. En las últimas pruebas realizadas en unos 10 segundos, esto redujo en gran medida el tiempo total de cálculo. Conclusión De acuerdo con los requisitos del modelo del algoritmo diseñado, se ha demostrado la ayuda práctica mediante la construcción de un modelo computacional. El algoritmo puede optimizarse y seleccionarse según el modelo de cálculo en función de la realidad de la aplicación. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Correlation between salivary biomarkers and plaque-induced gingivitis exacerbated by pregnancy

Objetive: To evaluate the correlation between salivary biomarkers (the salivary antioxidant ability, salivary level of polyphenols, and other antioxidants) with plaque-induced gingivitis exacerbated by pregnancy in pregnant and nonpregnant women. Material and Methods: For this observational study, medical records, dental examinations, and analyses of saliva samples were carried out in pregnant and nonpregnant women. A p-value of <0.05 was considered significant. Results: The pregnant women (n =17) exhibited a lower antioxidant capacity (p-value=0.0041), higher levels of polyphenols, gingival index, bleeding on probing, and subjects consuming mineral-enriched products (p-value from <0.0001 to 0.0466), and unchanged levels of phosphotungstic acid reactive substances, proteins, oral hygienic habits, plaque index and probing depth (p-value from 0.0683 to 0.8358), in comparison with the nonpregnant women (n=9). Also, a positive correlation between the gingival index and salivary polyphenol content was observed (r-value = 0.4087, p-value = 0.0202). Conclusion: The salivary polyphenols correlate with plaque-induced gingivitis exacerbated by pregnancy, suggesting a deficiency of salivary antioxidant protection.

Objetivo: Evaluar la correlación entre los biomarcadores salivales (la capacidad antioxidante salival, el nivel salival de polifenoles y otros antioxidantes) con la gingivitis inducida por placa exacerbada por el embarazo en mujeres embarazadas y no embarazadas. Material y Métodos: Para este estudio observacional, se realizaron registros médicos, exámenes dentales y análisis de muestras de saliva en mujeres embarazadas y no embarazadas. Se consideró significativo un valor de p<0,05. Resultados: Las gestantes (n=17) presentaron menor capacidad antioxidante (p=0,0041), mayores niveles de polifenoles, índice gingival, sangrado al sondaje y los sujetos que consumían productos enriquecidos con minerales (p<0,0001 a p<0,0466), y no hubo diferencias en los niveles de sustancias reactivas al ácido fosfotúngstico, proteínas, hábitos de higiene bucal, índice de placa y profundidad de sondaje (p=0,0683 a 0,8358), en comparación con las mujeres no embarazadas (n=19). Además, se observó una correlación positiva entre el índice gingival y elcontenido de polifenoles salivales (r = 0,4087, p= 0,0202). Conclusión: Los polifenoles salivales se correlacionan con la gingivitis inducida por placa y exacerbada por el embarazo, lo que sugiere una deficiencia de protección antioxidante salival.

Utilidad diagnóstica del dímero-D en el tromboembolismo pulmonar de pacientes COVID-19 / DiagnosticutilityofD-dimerin pulmonarythromboembolism in COVID-19 patients

Resumen Introducción: El aumento de la concentración de dímero-D en pacientes COVID-19 se ha asociado a mayor gravedad y peor pronóstico; sin embargo, su rol en predecir el diagnóstico de tromboembolismo pulmonar (TEP), aún es incierto. Objetivo: Evaluar la utilidad del dímero-D plasmático en el diagnóstico de TEP en pacientes con COVID-19. Pacientes y Métodos: Estudio observacional analítico. Se incluyó a pacientes COVID-19 que tenían una angiotomografía computada de tórax (AngioTAC). Se registraron datos clínicos, niveles plasmáticos de dímero-D de ingreso y previo al momento de realizar la AngioTAC. Se identificó la presencia o ausencia de TEP. Resultados: Se incluyeron 163 pacientes; 37(23%) presentaron TEP. Al comparar la serie de pacientes con TEP versus sin TEP, no se encontraron diferencias significativas en características clínicas, ni mortalidad. Hubo diferencias significativas en el nivel plasmático del dímero-D previo a realizar la AngioTAC (3.929 versus 1.912 μg/L; p = 0,005). El área bajo la curva ROC del dímero-D para TEPfue de 0,65. El mejor punto de corte del dímero-D fue de 2.000 μg/L, con una baja sensibilidad y valor predictivo positivo. El valor de corte con el mejor valor predictivo negativo (VPN)fue de 900 μg/L (96%), el cual fue mejor que la estrategia de corte de dímero D ajustado por edad (VPN 90%). Conclusión: La capacidad discriminativa del dímero D para diagnosticar TEP fue baja. En cambio, el dímero D mantiene un alto valor predictivo negativo para descartar TEP, el cual es mayor al valor descrito clásicamente en los pacientes no COVID.

Introduction: Increased D-dimer concentration in COVID-19 patients has been associated with greater severity and worse prognosis; however its role in predicting the diagnosis of pulmonary thromboembolism (PTE), is still uncertain. Objective: To evaluate the usefulness of plasma D-dimer in the diagnosis of PTE in patients with COVID-19. Method: Analytical observational study. COVID-19 patients who had a chest computed tomography angiography (CTA) were included. Clinical data, Ddimer plasma levels on admission and prior to CTA were recorded. The presence or absence of PTE was identified. Results: 163 patients were included, 37 (23%) presented PTE. After comparing the series of patients with PTE versus the series without PTE, no significant differences were found in clinical characteristics or mortality. There were significant differences in the plasma level of D-dimer prior to performing CTA (3,929 μg/L versus. 1,912 μg/L; p = 0.005). The area under the D-dimer ROC curve for PTEprediction was 0.65. The best D-dimer cutoffpoint was 2.000μg/L, with a low sensitivity and positivepredictive value. The cutoff value with the best negativepredictive value (NPV) was 900 μg/L (96%), which was better than the age-adjusted D-dimer cutoff strategy (NPV 90%). Conclusion: The discriminative ability of D-dimer to diagnose PTE was low. In contrast, D-dimer maintains a high negative predictive value to rule out PTE, which is higher than the value classically described in non-COVID patients.

Gingival crevicular fluid as Biomarker's source for alzheimer's disease / Fluido crevicular gingival como biomarcador de enfermedad de Alzheimer

Abstract Periodontitis is a low-grade inflammatory disease caused by a subgingival dysbiotic microbiota. Multiple studies have determined the higher prevalence of tooth loss and poor oral hygiene in patients with Alzheimer's disease (AD). However, the periodontal diagnosis, periodontal bacteria or mediators has not been measured to date. Aim: To determine the periodontal status, the pro-inflammatory mediators, Porphyromonas gingivalis load, and Apoliporpotein E (ApoE) in patients with AD. A complete dental examination was performed on 30 patients, and cognitive status was determined by the Montreal Cognitive Assessment (MoCA). Subgingival microbiota and GCF samples were then taken from all patients from the deepest sites. Total DNA was isolated from the microbiota samples for the quantification of the 16S ribosomal subunit. Pro-inflammatory mediators and ApoE were quantified from the gingival crevicular fluid (GCF). Patients with AD had periodontitis stage III-IV in 80%, a higher concentration of pro-inflammatory and ApoE mediators, and a higher P. gingivalis load compared to healthy subjects. The pro-inflammatory mediators, P. gingivalis load had a negative correlation with the MoCA test scores. Finally, a ROC curve was performed to assess the specificity and sensitivity of ApoE levels, detecting an area of 84.9%. In AD patients, we found a more severe periodontitis, a higher levels of pro-inflammatory mediators, and higher bacterial load. In addition, there is an increase in ApoE that allows to clearly determine patients with health, periodontitis and periodontitis and AD.

Resumen La periodontitis es una enfermedad crónica no transmisible que se caracteriza por generar una inflamación sistémica de bajo grado causada por una microbiota disbiótica subgingival. Múltiples estudios han determinado la mayor prevalencia de pérdida de dientes y mala higiene bucal en pacientes con enfermedad de Alzheimer (EA). Sin embargo, el diagnóstico periodontal, bacterias periodontales o mediadores pro-inflamatorio no se ha medido hasta la fecha. Determinar el estado periodontal, los mediadores pro-inflamatorios, la carga de Porphyromonas gingivalis y la apoliporpoteína E (ApoE) en pacientes con EA. Se realizó un examen odontológico completo en 30 pacientes y el estado cognitivo se determinó mediante la Evaluación Cognitiva de Montreal (MoCA). Luego, se tomaron muestras de microbiota subgingival y FCG de todos los pacientes de los sitios más profundos. Se aisló el DNA total de las muestras de microbiota para la cuantificación de la subunidad ribosómica 16S. Los mediadores pro-inflamatorios y la ApoE se cuantificaron a partir del líquido crevicular gingival (GCF). Los pacientes con EA tenían periodontitis en estadio III-IV en 80%, una mayor concentración de mediadores pro-inflamatorios y ApoE, y una mayor carga de P. gingivalis en comparación con los sujetos sanos. Los mediadores pro-inflamatorios y la carga de P. gingivalis tuvieron una correlación negativa con las puntuaciones de la prueba MoCA. Finalmente, se realizó una curva ROC para evaluar la especificidad y sensibilidad de los niveles de ApoE, detectando un área del 84,9%. En los pacientes con EA encontramos una periodontitis más severa, mayores niveles de mediadores pro-inflamatorios y mayor carga bacteriana. Además, un aumento de ApoE que permite determinar claramente a los pacientes con salud, periodontitis y periodontitis y EA.

Manifestaciones pulmonares al diagnóstico de 170 pacientes con vasculitis asociadas a anticuerpos anticitoplasma de neutrófilos: cohorte retrospectiva de un centro universitario / Pulmonary manifestations at the diagnosis of 170 patients with vasculitis associated with anticytoplasmic neutrophil antibodies: retrospective cohort of a university center

INTRODUCCIÓN: Las Vasculitis Asociadas a Anticuerpos Anticitoplasma de Neutrófilos (VAA) son infrecuentes, pero de amplio espectro de presentación. Si bien el consenso de clasificación de Chapel Hill del año 2012, sigue vigente, la tendencia actual es clasificarlas de acuerdo al marcador inmunológico: anti-Proteinasa 3 (PR3) o anti-mieloperoxidasa (MPO). Las manifestaciones pulmonares clásicas son la hemorragia alveolar y los nódulos pulmonares. En los últimos 10 años se ha descrito la enfermedad pulmonar difusa (EPD). Los estudios epidemiológicos son escasos, y suelen representar en su mayoría poblaciones norteamericanas o europeas. El objetivo es describir las características del compromiso pulmonar al debut en VAA en un centro universitario. PACIENTES Y MÉTODO: De un total de 190 pacientes con diagnóstico de VAA se incluyeron 170 en seguimiento en nuestro centro. Se revisaron aspectos clínicos, demográficos, laboratorio e imagenológicos de los pacientes incluidos. RESULTADOS: De los 170 pacientes, 112 (65,88%) presentaron compromiso pulmonar. 106 (94,64%) de los pacientes fueron anticuerpos anti citoplasma de neutrófilos (ANCA) positivos; de estos, 56 (53,27%) MPO (+) y 39 (36,45%) PR-3 (+). Un tercio de los pacientes de ambos grupos presentó hemorragia alveolar. En los pacientes MPO (+) predomina la EPD (53,5%) y en PR-3 (+) los nódulos pulmonares (69,23%). Destaca la baja frecuencia de patología obstructiva asociada. CONCLUSIONES: El compromiso pulmonar en las VAA es prevalente y heterogéneo. En nuestra serie, destaca la frecuencia de EPD en VAA MPO (+), lo que releva la importancia del estudio con ANCA en paciente con diagnóstico y seguimiento por EPD.

INTRODUCTION: Antineutrophil Cytoplasmic Antibodies (ANCA) associated vasculitis (AAV) are uncommon, but of broad spectrum of presentation. Although the 2012 Chapel Hill classification consensus remains valid, the current trend is to classify them according to the immunological marker: anti-Proteinase 3 antibody (PR-3) or anti-Myeloperoxidase antibody (MPO). The classic pulmonary manifestations are alveolar hemorrhage and pulmonary nodules. Interstitial lung disease (ILD) has been described in the last 10 years. Epidemiological studies are scarce, and they usually represent mostly North American or European populations. The objective is to describe the characteristics of lung involvement upon debut in AAV in a university center. PATIENTS AND METHODS: Of a total of 190 patients diagnosed with AAV, 170 were included in follow-up at our center. Clinical, demographic, laboratory and imaging aspects of the included patients were reviewed. RESULTS: Of the 170 patients, 112 (65.88%) had lung involvement. 106 (94.64%) of the patients were ANCA (+); of these, 56 (53.27%) MPO (+) and 39 (36.45%) PR-3 (+). One third of the patients in both groups had alveolar hemorrhage. In MPO (+) patients, ILD predominates (53.5%) and in PR-3 (+) pulmonary nodules (69.23%). The low frequency of associated obstructive pathology stands out. CONCLUSIONS: Pulmonary manifestations in AAVs are frequent and heterogeneous. Locally, the association of ILD and AAV MPO (+) stands out, which highlights the importance of ANCA study in patients with diagnosis and follow-up by ILD.

Quality markers of Zingiberis Rhizoma Carbonisata before and after processing / 中国中药杂志

Based on the previous research results of our group and literature research, the chemical components, mechanisms, pharmacodynamics, and pharmacokinetics of Zingiberis Rhizoma Carbonisata were summarized to determine the quality markers(Q-markers) of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Our research group has clarified the differential components of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma, the meridian-warming hemostatic effect of Zingiberis Rhizoma Carbonisata, the related targets and pathways of the effect, the endogenous biomarkers of Zingiberis Rhizoma Carbonisata, and the hemodynamic processes of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Moreover, based on high-performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry(HPLC-DAD-ESIMS), a method for determining the content of Q-mar-kers was established. In conclusion, the study finally determined that gingerone, 6-shogaol, and diacetyl-6-gingerol were the Q-mar-kers of Zingiberis Rhizoma Carbonisata decoction pieces, and 6-gingerol, 8-gingerol, and 10-gingerol were Q-markers of Zingiberis Rhizoma decoction pieces. The result is expected to provide a reference for the establishment of quality standards for Zingiberis Rhizoma Carbonisata decoction pieces and Zingiberis Rhizoma decoction pieces.

Assessment of carbamazepine acute toxicity in the cockle Cerastoderma edule through chemical, physiological and biochemical tools / Avaliação da toxicidade aguda da carbamazepina no berbigão Cerastoderma edule por meio de ferramentas químicas, fisiológicas e bioquímicas

The cockle Cerastoderma edule was exposed to four concentrations (5, 10, 20 and 70 μg L-¹) of carbamazepine (CBZ). This anticonvulsant was found to alter the mussel behavior of by reducing its clearance rate (CR). Analysis of CBZ accumulation in tissues of C. edule was carried out using HPLC-UV after 48 or 96 hours of exposure. In addition, an overproduction of H2O2 by the bivalves was detected following exposure to CBZ but nitrite levels remained unchanged. Moreover, superoxide dismutase and catalase activities showed a significant increase in relation to their contact with CBZ. The activity of the biotransformation enzyme gluthatione-S-transferase did not change during exposure. Malondialdehyde (MDA) levels indicating cellular damage, increased when bivalves were exposed to 20 and 70 μg l-¹ of carbamazepine for 96 h CBZ. The results also indicate that acetylcholinesterase activity (AChE) was inhibited in all CBZ concentrations during the 48 h exposure period. However, during the 96 h exposure period, AChE was only inhibited at the highest concentration. Further studies are needed now for more exploration of the toxicity of CBZ since it could be bioaccumulable throughout the food web and may affect non-target organisms.

O berbigão Cerastoderma edule foi exposto a quatro concentrações (5, 10, 20 e 70 μg L-¹) de carbamazepina (CBZ). Este anticonvulsivante alterou o comportamento do mexilhão, reduzindo sua taxa de depuração (CR). A análise do acúmulo de CBZ nos tecidos de C. edule foi realizada por HPLC-UV após 48 ou 96 horas de exposição. Além disso, uma superprodução de H2O2 pelos bivalves foi detectada após a exposição à CBZ, mas os níveis de nitrito permaneceram inalterados. Além disso, as atividades de superóxido dismutase e catalase apresentaram aumento significativo em relação ao contato com CBZ. A atividade da enzima de biotransformação glutationa-S-transferase não se alterou durante a exposição. Os níveis de malondialdeído (MDA), indicando dano celular, aumentaram quando os bivalves foram expostos a 20 e 70 μg l-1 de carbamazepina por 96 h CBZ. Os resultados também indicam que a atividade da acetilcolinesterase (AChE) foi inibida em todas as concentrações de CBZ durante o período de exposição de 48 horas. No entanto, durante o período de exposição de 96 horas, a AChE foi inibida apenas na concentração mais alta. Mais estudos são necessários agora para uma maior exploração da toxicidade da CBZ, uma vez que pode ser bioacumulável em toda a cadeia alimentar e pode afetar organismos não alvo.

Effect of trans-resveratrol on oxidative stress biomarkers associated with atherosclerosis / Efeito do trans-resveratrol em biomarcadores de estresse oxidativo associados à aterosclerose

Growing evidence indicates that oxidative stress plays an important role in the pathophysiology of many cardiovascular diseases, including atherosclerosis. In this context, the use of bioactive compounds with antioxidant action can bring health benefits, especially in the prevention and control of pathophysiological events. Studies suggest that the polyphenol trans-resveratrol can reduce oxidative stress by acting on the nuclear factor erythroid-2-related factor 2 (Nrf2) and this effect would be associated with dosage. Thus, the present study aimed to investigate the effect of different doses of trans-resveratrol on biomarkers related to atherosclerosis and oxidative stress. In the first step, 27 randomized clinical trials, which evaluated the effect of trans-resveratrol on atherosclerosis-related biomarkers, were classified according to their protocol characteristics and profile of each outcome. Biochemical data from 12 biomarkers were selected to calculate the net change (%). Using multivariate analysis, the trials were distributed into 3 clusters. The studies that composed Clusters II and III were more effective in improving blood pressure and reducing dyslipidemia, respectively. These studies were characterized by a longer intervention time (> 2 months) with doses of around 200-500 mg/day. These results showed that the effects of transresveratrol are mainly related to dosage and intervention time. Based on these results, two doses were selected to apply in an experimental protocol to investigate the effect of trans-resveratrol on hepatic oxidative stress biomarkers mediated by Nrf2 pathway. LDLr(-/-) mice were fed for 8 weeks on a standard diet, followed by over 24 weeks on a Western diet, both containing trans-resveratrol at doses of 250 mg/kg diet/day (low dose resveratrol, LRD) or 400 mg/kg diet/day (high dose resveratrol, HRD). A control group (CONT) was maintained without supplementation. In general, both doses of trans-resveratrol did not affect the body weight and lipid profile of the animals. Only the LRD group showed reduced levels of two important biomarkers of oxidative stress in the liver (GSH/GSSG ratio and malonaldehyde), besides to reduced expression of factor nuclear kappa B (NF-kB). However, contrary to our hypothesis, both doses reduced Nrf2 expression in the liver compared to the CONT group. Regarding inflammatory cytokines, no changes were observed in the levels of TNF-α and IL-10. Furthermore, both doses increased the level of the pro-inflammatory cytokine IL-6. Taken together, our results suggest that trans-resveratrol supplementation at doses lower than 500 mg/day may contribute to the reduction of biomarkers related to atherosclerosis and oxidative stress

Evidências crescentes indicam que o estresse oxidativo desempenha um papel importante na fisiopatologia de muitas doenças cardiovasculares, incluindo a aterosclerose. Nesse contexto, o uso de compostos bioativos com ação antioxidante pode trazer benefícios à saúde, principalmente na prevenção e controle de eventos fisiopatológicos. Estudos sugerem que o polifenol trans-resveratrol pode reduzir o estresse oxidativo atuando na via do fator nuclear eritroide 2 relacionado ao fator 2 (Nrf2) e que esse efeito estaria associado a dosagem. Assim, o presente estudo teve como objetivo investigar o efeito de diferentes doses de trans-resveratrol sobre biomarcadores relacionados à aterosclerose e estresse oxidativo. Na primeira etapa, 27 ensaios clínicos randomizados, que avaliaram o efeito do trans-resveratrol em biomarcadores relacionados à aterosclerose, foram classificados de acordo com suas características de protocolo e perfil de cada resultado. Dados bioquímicos de 12 biomarcadores foram selecionados para calcular a variação líquida (%). Usando análise multivariada, os ensaios foram distribuídos em 3 Clusters. Os estudos que compuseram os Clusters II e III foram mais eficazes na melhora da pressão arterial e na redução da dislipidemia, respectivamente. Esses estudos foram caracterizados por um tempo de intervenção mais longo (> 2 meses) com doses de cerca de 200-500 mg/dia. Esses resultados mostraram que os efeitos do transresveratrol estão relacionados principalmente à dosagem e ao tempo de intervenção. Com base nesses resultados, duas doses foram selecionadas para aplicar em um protocolo experimental para investigar o efeito do trans-resveratrol em biomarcadores de estresse oxidativo hepático mediados pela via do Nrf2. Camundongos LDLr(-/-) foram alimentados por 8 semanas com dieta padrão, seguidos por mais de 24 semanas com Western diet, ambos contendo trans-resveratrol nas doses de 250 mg/kg de dieta/dia (baixa dose de resveratrol, LRD) ou 400 mg/kg de dieta/dia (alta dose de resveratrol, HRD). Um grupo controle (CONT) foi mantido sem suplementação. Em geral, ambas as doses de trans-resveratrol não afetaram o peso corporal e o perfil lipídico dos animais. Apenas o grupo LRD apresentou níveis reduzidos de dois importantes biomarcadores de estresse oxidativo no fígado (razão GSH/GSSG e malonaldeído), além da redução da expressão de fator nuclear kappa B (NF-kB). No entanto, ao contrário da nossa hipótese, ambas as doses reduziram a expressão de Nrf2 no fígado em comparação com o grupo CONT. Em relação às citocinas inflamatórias, não foram observadas alterações nos níveis de TNF-α e IL-10. Além disso, ambas as doses aumentaram o nível da citocina pró-inflamatória IL-6. Em conjunto, nossos resultados sugerem que a suplementação de trans-resveratrol em doses menores de 500 mg/dia podem contribuir para a redução de biomarcadores relacionados à aterosclerose e ao estresse oxidativo

Identification of new human mesenchymal stem cell biomarker by aptamers / Identificação de novos biomarcadores de células tronco mesenquimais de origem humana por meio de aptâmeros

Stem cells are undifferentiated cells that can be distinguished from others by their ability to self-renew and to differentiate into new specific cell types. Mesenchymal stem cells (MSC) are adult stem cells that can be obtained from different sources, such as adipose tissue, bone marrow, dental pulp, and umbilical cord. They can either replicate, originating new identical cells, or differentiate into cells of mesodermal origin and from other germ layers. MSC have been studied as new tools for regenerative therapy. Although encouraging results have been demonstrated, MSC-based therapies still face a great barrier: the difficulty of isolating these cells from heterogeneous environments. MSC are currently characterized by immunolabelling through a set of multiple surface membrane markers, including CD29, CD73, CD90 and CD105, which are also expressed by other cell types. Hence, the present work aimed to identify new specific biomarkers for the characterization of human MSC using DNA aptamers produced by the SELEX (Systematic Evolution of Ligands by EXponential Enrichment) technique. Our results showed that MSC from different origins bound to DNA candidate aptamers, that is, DNA or RNA oligonucleotides selected from random libraries that bind specifically to biological targets. Aptamer-bound MSC could be isolated by fluorescenceactivated cell sorting (FACS) procedures, enhancing the induction of differentiation into specific phenotypes (chondrocytes, osteocytes and adipocytes) when compared to the whole MSC population. Flow cytometry analyses revealed that candidate aptamers bound to 50% of the MSC population from dental pulp and did not present significant binding rates to human fibroblasts or lymphocytes, both used as negative control. Moreover, immunofluorescence images and confocal analyses revealed staining of MSC by aptamers localized in the surfacemembrane of these cells. The results also showed internal staining of human monocytes by our investigated aptamers. A non-specific control aptamer (CNTR APT) obtained from the random pool was then utilized to compare the specificity of the aptamers bound to the analyzed non-apoptotic cells, showing no staining for MSC. However, 40% of the monocytes bound to the CNTR APT. Normalized data based on the cells bound to candidate aptamers compared to those bound to the CNTR APT, revealed a 10 to 16-fold higher binding rate for MSC against 2-fold for monocytes. Despite its low specificity, monocyte-aptamer binding occurs probably due to the expression of shared markers with MSC, since monocytes are derived from hematopoietic stem cells and are important for the immune system ability to internalize/phagocyte external molecules. Given that, we performed a pull-down assay followed by mass spectrometry analysis to detect which MSC-specific protein or other target epitope not coexpressed by monocytes or the CNTR APT would bind to the candidate aptamer. Distinguishing between MSC and monocyte epitopes is important, as both cells are involved in immunomodulatory effects after MSC transplantations. ADAM17 was found to be a target of the APT10, emerging as a possible biomarker of MSC, since its involvement in the inhibition of the TGF signaling cascade, which is responsible for the differentiation of MSC. Thus, MSC with a higher stemness profile should overexpress the protein ADAM17, which presents a catalytic site with affinity to APT10. Another target of Apt 10 is VAMP3, belonging to a transmembrane protein complex that is involved in endocytosis and exocytosis processes during immune and inflammatory responses. Overall, proteins identified as targets of APT10 may be cell surface MSC biomarkers, with importance for MSC-based cell and immune therapies

Células tronco são células indiferenciadas que podem ser distinguidas de outros tipos celulares por meio da habilidade de se auto renovarem e de se diferenciarem em novos tipos celulares. Células tronco mesenquimais (MSC) são células tronco adultas encontradas em diferentes tecidos como tecido adiposo, polpa de dente e cordão umbilical. Estas células podem se autodividir em células idênticas ou se diferenciarem em células de origem mesodermal. Estas células têm sido estudadas em novas aplicações que envolvem terapia regenerativas. Embora resultados encorajadores tenham sido demonstrados, terapias que utilizam MSC ainda encontram uma grande barreira: a dificuldade no isolamento destas células a partir de um ambiente heterogêneo. MSC são caracterizadas por populações positivas em ensaios de imunomarcação para os epítopos membranares CD29, CD73, CD90 e CD105, presentes também em outros tipos celulares. Assim, o presente trabalho tem o objetivo de identificar novos biomarcadores de MSC de origem humana, utilizando aptâmeros de DNA produzidos pela técnica SELEX (Systematic Evolution of Ligands by EXponential Enrichment) como ferramenta. Nossos resultados mostraram que MSC de diferentes origens ligam-se a aptâmeros (oligonucleotídeos de DNA ou RNA que atuam como ligantes específicos de alvos moleculares) de DNA candidatos que atuam no isolamento de MSC por meio da técnica FACS de separação celular, promovendo uma maior indução de diferenciação em células específicas (condrócitos, osteócitos e adipócitos) comparada com a população total de MSC. Análises de citometria de fluxo mostraram que os aptâmeros candidatos se ligam a 50% das MSC de polpa de dente e não apresentam taxa de ligação significante para fibroblastos e linfócitos de origem humana - utilizados como controles negativo. Além domais, imagens de imunofluorescência e confocal mostraram ligação na superfície da membrana de MSC e a marcação interna de monócitos a estes aptâmeros. Portanto, um aptâmero controle (CNTR APT) foi utilizado para comparar a especificidade dos aptâmeros ligados a células viáveis, mostrando a não ligação deste aptâmero a MSC. Porém, 40% da população de monócitos ligou-se ao CNTR APT. Uma normalização baseada na comparação entre as taxas de ligação entre células ligadas com aptâmeros candidatos e o aptâmero controle gerou uma taxa de especificidade entre 10-16 vezes maior para MSC contra 2,5 vezes para os monócitos. Deste modo, embora os resultados tenham mostrado uma taxa de ligação entre monócitos e aptâmeros, as MSC ligadas aos aptâmeros candidatos possuem uma maior taxa de especificidade devido a uma maior presença de antígenos que são expressos em ambas as células. Um ensaio de Pull Down seguido de espectrometria de massas foi utilizado para a identificação de biomarcadores que se ligariam aos aptâmeros candidatos, e que não seriam co-expressos por monócitos e por antígenos ligados ao aptâmero controle. Deste modo, a proteína ADAM17 foi identificada nas amostras de APT10 ligadas às MSC. Tal proteína está relacionada à inibição de uma cascata de sinalização da família de proteínas TGF, responsável pela diferenciação de MSC. Assim, MSC com maior potencial tronco deveriam expressar ADAM17 em maior quantidade. Tal proteína apresenta um sítio catalítico que demonstra interagir com o APT10, de acordo com predição Docking entre proteína e DNA. Foi identificada também, a proteína VAMP3, que pertence a um complexo proteico transmembranar responsável pelos processos de endocitose e exocitose, e que podem ter um papel importante na liberação de citocinas e outras moléculas relacionadas às respostas imune e inflamatórias. Deste modo, o APT10 identificou proteínas importantes que devem estar relacionas com a melhora de imunoterapias que utilizam MSC

Strain longitudinal global como preditor de cardiotoxicidade induzida por quimioterapia / Global longitudinal strain as predictor of chemotherapy-induced cardiotoxicity

Fundamento: A cardiotoxicidade induzida por quimioterapia (CiC) é uma complicação importante entre os pacientes que recebem antraciclinas. Biomarcadores e parâmetros de imagem têm sido estudados por sua capacidade de identificar pacientes com risco de desenvolver essa complicação. O strain longitudinal global do ventrículo esquerdo (SLG-VE) tem sido descrito como um parâmetro sensível para detectar disfunção sistólica, mesmo na presença de fração de ejeção do ventrículo esquerdo (FEVE) preservada. Objetivo: avaliar o papel do SLG-VE como preditor de CiC. Métodos: O presente estudo consiste em uma análise post-hoc do estudo CECCY (Carvedilol for Prevention of ChemotherapyRelated Cardiotoxicity [Carvedilol para Prevenção da Cardiotoxicidade Relacionada à Quimioterapia]), que avaliou a prevenção primária de cardiotoxicidade com carvedilol durante quimioterapia com doxorrubicina em uma população com câncer de mama. Definiu-se cardiotoxicidade como uma redução >10% na FEVE. O SLG-VE foi obtido antes da quimioterapia em pacientes sem doença cardiovascular prévia ou anormalidades no ecocardiograma. Resultados: Trinta e um pacientes submetidos a estudo ecocardiográfico completo incluindo avaliação de SLG-VE antes da quimioterapia foram incluídos nesta análise. Um SLG-VE absoluto <16,9% antes da quimioterapia mostrou 100% de sensibilidade e 73% de especificidade para predizer cardiotoxicidade (AUC=0,85; IC 95% 0,680­0,959, p<0,001). Nesta população, os valores de FEVE antes da quimioterapia não foram preditores de CiC (IC 95% 0,478 a -0,842, p=0,17). A associação de baixos níveis séricos de SLG-VE (<17%) e BNP (>17 pg/mL) dois meses após a quimioterapia aumentou a precisão para detectar CiC de início precoce (100% de sensibilidade, 88% de especificidade, AUC=0,94; IC 95% 0,7810,995, p<0,0001). Conclusões: Nossos dados sugerem que o SLG-VE é um possível preditor de cardiotoxicidade induzida por quimioterapia. São necessários estudos maiores para confirmar a relevância clínica desse parâmetro ecocardiográfico nesse cenário clínico. (AU)

Background: Chemotherapy-induced cardiotoxicity (ChC) is an important complication among patients receiving anthracyclines. Biomarkers and imaging parameters have been studied for their ability to identify patients at risk of developing ChC. Left ventricular global longitudinal strain (LV-GLS) is a sensitive parameter for detecting systolic dysfunction despite the presence of preserved left ventricular ejection fraction (LVEF). Objective: To evaluate the role of the LV-GLS as a predictor of ChC. Methods: This was a post-hoc analysis of the Carvedilol for Prevention of Chemotherapy-Related Cardiotoxicity trial, which evaluated the primary prevention of cardiotoxicity with carvedilol during doxorubicin chemotherapy in a population of patients with breast cancer. Cardiotoxicity was defined as a reduction ≥10% in LVEF. LV-GLS was determined before chemotherapy in patients with no prior cardiovascular disease or echocardiogram abnormalities. Results: Thirty-one patients for whom a complete echocardiography study including measurement of LV-GLS was performed before chemotherapy were included in this analysis. An absolute LV-GLS<16.9% before chemotherapy showed 100% sensitivity and 73% specificity for predicting cardiotoxicity (area under the curve [AUC], 0.85; 95% confidence interval [CI], 0.680­0.959; p<0.001). In this population, LVEF values before chemotherapy did not predict ChC (95% CI, 0.478 to -0.842; p=0.17). The association of low LV-GLS (<17%) and brain-type natriuretic peptide serum levels (>17 pg/mL) at 2 months after chemotherapy increased the accuracy for detecting early-onset ChC (100% sensitivity, 88% specificity; AUC, 0.94; 95% CI, 0.781­0.995; p<0.0001). Conclusions: Our data suggest that LV-GLS is a potential predictor of ChC. Larger studies are needed to confirm its clinical relevance in this clinical setting. (AU)