Resumen (Objetivo) Tryapanosoma cruzi, un agente causal de la enfermedad de Chagas, es un parásito que presenta una alternancia en su ciclo de vida entre un hospedador invertebrado (triatomino) y uno vertebrado (mamífero). Varios estudios han demostrado que en T. cruzi, HSP83 (homólogo de HSP90) es esencial para la división celular y el control de la respuesta al estrés térmico. Esta investigación se concentró en el estudio de la clonación, la caracterización bioinformática y la expresión del gen de la proteína de choque térmico HSP83 de T. cruzi para estudios posteriores sobre señalización. (Metodología) El ARN fue extraído de epimagostigotes T. cruci (clon EPm6/MOHM/VE/2007/ 6c), utilizando un kit comercial. El ADNc que codifica HSP83 se obtuvo utilizando RT-PCR, a partir del ARNm extraído, para lo cual se diseñaron los cebadores con base en la secuencia HSP83 de la cepa T. cruzi CL Brener. La clonación se realizó usando pGEM®T-Easy y se subclonó en el vector de expresión pQE30. Se efectuó la caracterización bioinformática y de secuencias. El gen se expresó y la proteína recombinante se purificó por cromatografía de afinidad, igual que se identificó por inmunotransferencia. (Resultados) El análisis de secuencia mostró similitud con el gen HSP83 de Trypanosoma cruzi y se observaron dominios HSP, así como epítopos B en la secuencia. Después de 3 horas de inducción con IPTG, se obtuvo una proteína recombinante con un peso aproximado de 83 kDa. La reacción de inmunotransferencia con suero hiperinmune anti-epimastigote de T. cruzi permitió la detección de una sola banda con un peso molecular de aproximadamente 83 kDa. (Conclusiones) Todos los resultados indican que se logró la clonación y caracterización de HSP83 de Trypanosoma cruzi.

Resumo (Objetivo) O Tryapanosoma cruzi, agente causador da doença de Chagas, é um parasita que apresenta uma alternância em seu ciclo de vida entre um hospedeiro invertebrado triatomíneo) e um vertebrado(mamífero). Vários estudos demonstraram que, no T. cruzi, a HSP83 (homóloga da HSP90) é essencial para a divisão celular e o controle da resposta ao estresse térmico. Esta pesquisa concentrou-se no estudo da clonagem, caracterização bioinformática e expressão do gene da proteína de choque térmico HSP83 do T. cruzi para estudos adicionais de sinalização. (Metodologia) O RNA foi extraído de epimastigotas de T. cruzi (clone EPm6/MOHM/VE/2007/ 6c) usando um kit comercial. O cDNA que codifica a HSP83 foi obtido por meio de RT-PCR a partir do mRNA extraído, para o qual os primers foram projetados com base na sequência da HSP83 da cepa T. cruzi CL Brener. A clonagem foi realizada com o pGEM®T-Easy e subclonada no vetor de expressão pQE30. Foi realizada a caracterização bioinformática e de sequência. O gene foi expresso e a proteína recombinante foi purificada por cromatografia de afinidade e identificada por imunotransferência. (Resultados) A análise da sequência mostrou similaridade com o gene HSP83 do Trypanosoma cruzi e os domínios HSP, bem como os epítopos B, foram observados na sequência. Após 3 horas de indução com IPTG, foi obtida uma proteína recombinante com peso aproximado de 83 kDa. A imunotransferência com soro hiperimune antiepimastigota de T. cruzi permitiu a detecção de uma única banda com peso molecular de aproximadamente 83 kDa. (Conclusões) Todos os resultados indicam que a clonagem e a caracterização da HSP83 do Trypanosoma cruzi foram alcançadas.

Abstract (Objective) Trypanosoma cruzi, a causal agent of Chagas’ disease, is a parasite whose life cycle alternates between an invertebrate (triatomine) and a vertebrate (mammal) host. Various studies have shown that in T. cruzi, HSP83 (a homolog of HSP90) is essential for cell division and control of the response to thermal stress. This investigation focused on studying the cloning, bioinformatic characterization, and expression of the heat shock protein HSP83 T. cruzi gene for further cell signaling studies. (Methodology) RNA was extracted from T. cruzi epimastigotes (EPm6 clone, MHOM/VE/2007/ 6c) using a commercial kit. The cDNA encoding HSP83 was determined using RT-PCR on extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed using pGEM®T-Easy and subcloned into the expression vector pQE30. Sequence and bioinformatic characterization were performed. The gene was expressed, and the recombinant protein was purified using affinity chromatography and identified through immunoblotting. (Results) Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi, and HSP domains and B epitopes in the sequence were also observed. After 3 hours of induction with IPTG, a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum helped detect a single band with a molecular weight of nearly 83 kDa. (Conclusions) All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved.

Abstract: The main objective of this study is to characterize the virus associated with faba bean (Vicia faba L.) crops in the Department of Nariño (Colombia). Faba bean is an important food source, but it faces challenges from viral diseases such as "pod mottle." This disease manifests with symptoms such as leaf chlorosis, dwarfism, and pod lesions. Viral characterization is conducted through seed transmission tests, mechanical transmission, RT-PCR, and transmission electron microscopy (TEM). The results show that the virus morphology consists of spherical and enveloped particles that range between 80-100nm in diameter. RT-PCR and sequencing of the nucleocápside gen (558bp) reveals the virus is Orthotospovirus tomatomaculae (Tomato spotted wilt virus) with a 97% homology. Seed transmission is 2% while mechanical transmission is 40%. This is the first report of an Orthotospovirus affecting faba bean in Nariño, posing a risk to food security.

Resumen: El principal objetivo de este estudio es caracterizar el virus asociado al cultivo de haba (Vicia faba L.) en el Departamento de Nariño (Colombia). El cultivo de haba es una fuente importante de alimento, pero enfrenta desafíos por enfermedades virales como la "viruela de la vaina". Esta enfermedad se manifiesta con síntomas como clorosis en las hojas, enanismo y lesiones en la vaina. La caracterización del virus se realiza mediante pruebas de transmisión por semilla, transmisión mecánica, RT-PCR y microscopía electrónica de transmisión (TEM). Los resultados muestran que la morfología del virus consiste en partículas esféricas y envueltas de entre 80-100nm en diámetro. RT-PCR y secuenciación del gen nucleocápside (558pb) revela que el virus es Orthotospovirus tomatomaculae (Tomato spotted wilt virus) con una homología de 97%. La transmisión por semilla es de 2% y mecánicamente del 40%. Este es el primer informe de un Orthotospovirus afectando el haba en Nariño, lo que representa un riesgo para la seguridad alimentaria.

ABSTRACT Hop stunt viroid (HSVd) is the causal agent of citrus cachexia, which can lead to significant crop losses. As part of a preventive management strategy, it is essential to use healthy plants from a certification program. This study aimed to evaluate the healthiness of 17 citrus cultivars against HSVd using both biological and molecular diagnostics. For biological diagnosis, Etrog citron (Citrus medica L.) clone Arizona 861 S1 was used as an indicator plant, incorporating negative and positive controls for HSVd infection. For molecular diagnosis, reverse transcription polymerase chain reaction (RT-PCR) technique was employed with specific primers for viroid detection. Results indicated that six months after inoculation, the positive control exhibited symptoms associated with viroid infection, such as necrosis in the petiole and stem, leaf epinasty, and necrosis in the midvein. In contrast, both the negative control and evaluated citrus cultivars showed no symptoms. These findings were supported by molecular analysis, which allowed the amplification of a viroid genome region only in the positive control. Both biological and molecular diagnosis confirmed the absence of HSVd in the citrus varieties assessed. This study will allow the evaluation of the citrus plant collections used in the foundation category as a source of buds for plant multiplication of registered and certified seeds for citrus propagation nurseries in Colombia.

RESUMEN El viroide del enanismo del lúpulo (HSVd) es el agente causal de la cachexia de los cítricos, que puede provocar pérdidas significativas en el cultivo. Como parte de una estrategia de manejo preventivo, es indispensable utilizar plantas sanas provenientes de un programa de certificación. Este estudio tuvo como objetivo evaluar la sanidad de 17 cultivares de cítricos frente a HSVd, utilizando diagnósticos biológico y molecular. Para el diagnóstico biológico, se empleó como planta indicadora cidro Etrog (Citrus medica L.) clon Arizona 861 S1, incluyendo controles negativos y positivos para la infección por HSVd. Para el diagnóstico molecular se utilizó la técnica de Reacción en Cadena de la Polimerasa con Transcriptasa Inversa (RT-PCR) con cebadores específicos para la detección del viroide. Los resultados indicaron que seis meses después de la inoculación, el control positivo mostró síntomas asociados con la infección por el viroide, como necrosis en el peciolo y tallo, epinastia foliar y necrosis en vena media. Por el contrario, tanto el control negativo como los cultivares de cítricos evaluados no presentaron síntomas. Estos resultados fueron corroborados por el análisis molecular, que permitieron amplificar una región del genoma del viroide, únicamente en el control positivo. Ambos diagnósticos biológico y molecular confirmaron la ausencia de HSVd en las variedades de cítricos evaluadas. Este estudio permitirá la evaluación de las colecciones de plantas de cítricos utilizadas en la categoría fundación como fuente de yemas para la multiplicación de semilla registrada y certificadapara los viveros productores de material de propagación de cítricos en Colombia.

Abstract Objective: To evaluate if the comorbidity and coinfections presented by SARS-CoV-2 infection vs. COVID-19 impact our Mexican children. Method: Prospective and observational study that included the 2020-2021 peak influenza season. All patients with a diagnosis of infection by SARS-CoV-2 vs. COVID-19 who were admitted to the Hospital Infantil de Mexico were analyzed. Real-time RT-PCR for SARS-CoV-2 was performed in all patients, determining E, RdRp and RP genes and protein N, as well as RT-PCR for detection of respiratory viruses. Results: The inclusion criteria were met by 163 patients. The group with the highest risk of becoming ill was adolescents (40.4%), followed by schoolchildren and preschoolers (21.4% and 19.6% of the cases, respectively). There were three cases with viral coinfection: two (1.2%) with parvovirus B-19 and one (0.6%) with herpes type I; another two (1.2%) showed bacterial coinfection. The main comorbidity were obesity, acute lymphoblastic leukemia and arterial hypertension. Regarding mortality, we only had four cases (2.4%). Conclusions: Obesity, cancer, hypertension, heart disease and diabetes are comorbidity present in our patients, as referred to in literature, but not coinfections. In our study, we did not have any associated mortality related to comorbidity.

Resumen Objetivo: Evaluar el impacto de la comorbilidad y de las coinfecciones presentadas por la infección por SARS-CoV-2 vs. COVID-19 en niños mexicanos. Método: Estudio prospectivo y observacional que comprendió la temporada alta de influenza 2020-2021, analizando todos los pacientes con diagnóstico de infección vs. enfermedad por SARS-CoV-2 vs. COVID-19 que ingresaron al Hospital Infantil de México. Se realizó en todos RT-PCR en tiempo real para SARS-CoV-2, determinando gen E, gen RdRp, gen RP y proteína N, y RT-PCR multiplex para detección de virus respiratorios. Resultados: Los criterios de inclusión los cumplieron 163 pacientes. El grupo con mayor riesgo de enfermar fueron los adolescentes (40.4%), seguidos de los escolares y preescolares (21.4% y 19.6% de los casos, respectivamente). Hubo tres casos con coinfección viral: dos (1.2%) con parvovirus B-19 y uno (0.6%) con herpes tipo I; hubo otros dos (1.2%) con coinfección bacteriana. La principal comorbilidad correspondió a obesidad, leucemia linfoblástica aguda e hipertensión arterial. En cuanto a mortalidad, solo hubo cuatro casos (2.4%). Conclusiones: Obesidad, cáncer, hipertensión, cardiopatías y diabetes constituyen la comorbilidad en nuestros pacientes, como se refiere en la literatura, no así las coinfecciones. En nuestro estudio no hubo casos de mortalidad relacionada con la comorbilidad.

RESUMEN Introducción: La rápida transmisión y los síntomas graves asociados con la infección aguda por el virus de la chikungunya (CHIKV) en niños lo convierten en un problema de salud altamente preocupante. Objetivo: Este estudio tuvo como objetivo describir las características de la infección aguda por el virus de la chikungunya en niños del Departamento de Caaguazú, Paraguay. Materiales y métodos: Un estudio retrospectivo fue realizado en el Departamento de Caaguazú, Paraguay, en 2023, con todos los pacientes que acudieron al Hospital Regional dentro de los 5 días posteriores al desarrollo de síntomas característicos de la infección aguda por CHIKV y que dieron positivo para el virus mediante RT-PCR. Pacientes con infecciones por Dengue o Zika fueron excluidos. Se recogieron datos sobre las características clínicas mediante un formulario normalizado de registro de casos y se creó un conjunto de datos electrónicos para su análisis. Resultados: Se incluyeron en el estudio un total de 461 niños. 51,6% eran mujeres. Los casos se dividieron en grupos según la edad pediátrica: lactantes (0-23 meses, n=88, 19,1 %), preescolares (2-5 años, n=115, 24,9%), escolares (6-11 años, n=163, 35.4%), and adolescentes (12-17 años and 11 meses, n=95, 20.6 %). La mialgia (64,6%) y la artralgia (63.7%) fueron más prevalentes en escolares y adolescentes. El vómito (89%), la cefalea (89,4%) y el dolor retroorbitario (95%) fueron más comunes en preescolares, escolares y adolescentes, mientras que la erupción cutánea (39,5%) y las petequias (18,5%) fueron más prevalentes en lactantes. Cuatro niños fallecieron durante el período de estudio. Conclusión: La epidemia de virus de la CHIKV de 2023 en Paraguay tuvo diferentes presentaciones clínicas dependiendo de la edad de los niños afectados, con resultados fatales ocurriendo en un pequeño porcentaje de casos.

ABSTRACT Introduction: The rapid transmission and severe symptoms associated with acute chikungunya virus (CHIKV) infection in children make it a highly concerning health issue. Objective: This study aimed to describe the characteristics of acute chikungunya virus infection in children from the Department of Caaguazú, Paraguay. Material and Methods: A retrospective observational study was conducted in the Department of Caaguazú, Paraguay, in 2023, with all patients who came to the Regional Hospital within 5 days of developing characteristic symptoms of acute CHIKV infection and tested positive for the virus by RT-PCR. Patients with Dengue or Zika infections were excluded. We collected data on clinical characteristics using a standardized case record form and created an electronic dataset for analysis. Results: A total of 461 children were included in the study. 51.6% were women. Cases were divided into groups based on pediatric age: infants (0-23 months, n=88, 19.1%), preschoolers (2-5 years, n=115, 24.9%), schoolchildren (6-11 years, n=163, 35.4%), and adolescents (12-17 years and 11 months, n=95, 20.6%). Schoolchildren and adolescents experienced a higher prevalence of myalgia (64.6%) and arthralgia (63.7%). Vomiting (89%), headache (89.4%), and retro-orbital pain (95%) were more common in preschoolers, schoolchildren, and adolescents, while rash (39.5%) and petechiae (18.5%) were more prevalent in infants. Four children died during the study period. Conclusion: The 2023 CHIKV virus epidemic in Paraguay had different clinical presentations depending on the age of the affected children, with fatal outcomes occurring in a small percentage of cases.

RESUMEN Introducción. El dengue es la enfermedad transmitida por mosquitos con mayor propagación mundial en los últimos años. Presenta un amplio espectro de manifestaciones clínicas y, en ocasiones, evoluciona a un estado crítico llamado dengue grave. Su tratamiento es de sostén. La información disponible acerca de las características clínicas, epidemiológicas y de laboratorio de la enfermedad en la población pediátrica es limitada. Objetivo. Describir la epidemiología y las manifestaciones clínicas y de laboratorio de la enfermedad. Población y métodos. Estudio descriptivo, observacional y retrospectivo. Incluyó pacientes entre 1 y 180 meses asistidos por dengue probable o confirmado en un hospital de niños, desde el 01 de enero de 2020 hasta el 31 de mayo de 2020. Resultados. Se incluyeron 85 pacientes por criterios microbiológicos de positividad o clínico-epidemiológicos. Veinticinco (29 %) confirmados por RT-PCR, todos serotipos DENV-1. La mediana de edad fue de 108 meses (rango intercuartílico: 84-144). Las principales manifestaciones clínicas fueron fiebre, cefalea y mialgias. Los hallazgos de laboratorio más importantes fueron leucopenia, trombocitopenia y elevación de transaminasas. Conclusión. El reconocimiento y la comprensión de las alteraciones clínicas y de laboratorio que se presentan durante la enfermedad pueden permitir un abordaje eficaz y contribuir a la reducción de cuadros clínicos más graves en los niños.

ABSTRACT Objective. To determine the effect of metformin (MET) treatment on adipogenic predisposition of bone marrow progenitor cells (BMPC), bone marrow adiposity and bone biomechanical properties. Materials and methods. 20 young adult male Wistar rats were sorted into four groups. Each of the groups received the following in drinking water: 100% water (C); 20% fructose (F); metformin 100 mg/kg wt/day (M); or fructose plus metformin (FM). After five weeks the animals were sacrificed. Both humeri were dissected to obtain BMPC, and both femurs were dissected to evaluate medullary adiposity (histomorphometry) and biomechanical properties (3-point bending). BMPC were cultured in vitro in adipogenic medium to evaluate RUNX2, PPAR-γ and RAGE expression by RT-PCR, lipase activity and triglyceride accumulation. Results. The fructose-rich diet (group F) caused an increase in both triglycerides in vitro, and medullary adiposity in vivo; being partially or totally prevented by co-treatment with metformin (group FM). No differences were found in femoral biomechanical tests in vivo, nor in lipase activity and RUNX2/PPAR-γ ratio in vitro. DRF increased RAGE expression in BMPC, being prevented by co-treatment with MET. Conclusions. Metabolic syndrome induced by a fructose-rich diet increases femoral medullary adiposity and, in part, the adipogenic predisposition of BMPC. In turn, this can be totally or partially prevented by oral co-treatment with MET.

RESUMEN Objetivo. Determinar el efecto de un tratamiento con metformina (MET) sobre la predisposición adipogénica de células progenitoras de médula ósea (CPMO), adiposidad de la médula ósea y propiedades biomecánicas óseas. Materiales y métodos. 20 ratas Wistar machos adultos jóvenes fueron separados en cuatro grupos, recibiendo en agua de bebida: 100% agua (C); 20% de fructosa (F); metformina 100 mg/kg peso/día (M); o fructosa más metformina (FM). Tras cinco semanas se sacrificaron los animales, se diseccionaron ambos húmeros para obtener CPMO, y ambos fémures para evaluar adiposidad medular (histomorfometría) y propiedades biomecánicas (flexión a 3 puntos). Las CPMO se cultivaron in vitro en medio adipogénico para evaluar expresión de RUNX2, PPAR-γ y RAGE por RT-PCR, actividad de lipasa y acumulación de triglicéridos. Resultados. La dieta rica en fructosa (grupo F) produjo un aumento tanto de triglicéridos in vitro, como de la adiposidad medular in vivo; siendo parcial o totalmente prevenido por un co-tratamiento con metformina (grupo FM). No se observaron diferencias en las pruebas biomecánicas femorales in vivo, ni en actividad de lipasa y relación RUNX2/PPAR-γ in vitro. La DRF aumentó la expresión de RAGE en CPMO, siendo prevenido por co-tratamiento con MET. Conclusiones. El síndrome metabólico inducido por una dieta rica en fructosa aumenta la adiposidad medular femoral y, en parte, la predisposición adipogénica de las CPMO. A su vez, esto puede ser prevenido total o parcialmente por un co-tratamiento oral con MET.

Resumo Kirenol (KNL) foi recentemente relatado como tendo propriedades anti-inflamatórias. No entanto, pouco se sabe sobre os potenciais mecanismos de suas propriedades anti-inflamatórias. Em HUVECs, elucidamos os mecanismos anti-inflamatórios do kirenol. RT-PCR foi usado para testar mRNA de mediadores pró-inflamatórios produzidos por Ox-LDL. A viabilidade das células foi medida usando MTT. Western blots analisaram os níveis de proteína. Em HUVECs estimuladas por Ox-LDL, o KNL inibiu significativamente a produção de mediadores pró-inflamatórios como NO, IL-1β, iNOS, TNF-α e IL-6. A expressão de p38, ROS e Nrf2 foi inibida por KNL. A inibição de p38, ROS e KNL causou acúmulo nuclear de Nrf2. O KNL também atenuou a fosforilação induzida por Ox-LDL de ERK1/2 e p38. Com base em nossos resultados, o KNL inibe a sinalização de NF-кB e MAPK em HUVECs, ativando a sinalização de Nrf2. Existe a possibilidade de que o KNL possa ser desenvolvido em um medicamento anti-inflamatório.

Abstract Kirenol (KNL) has recently been reported to have anti-inflammatory properties. Yet, little is known about the potential mechanisms of its anti-inflammatory properties. In HUVECs, we elucidated the anti-inflammatory mechanisms of kirenol. RT-PCR was used to test mRNA of pro-inflammatory mediators produced by Ox-LDL. The viability of cells was measured using MTT. Western blots analyzed protein levels. On Ox-LDL-stimulated HUVECs, KNL significantly inhibited the production of pro-inflammatory mediators such as NO, IL-1β, iNOS, TNF-α and IL-6. p38, ROS and Nrf2 expression were inhibited by KNL. Inhibition of p38, ROS, and KNL caused nuclear accumulation of Nrf2. KNL attenuated Ox-LDL-induced phosphorylation of ERK1/2 and p38, too. Based on our results, KNL inhibits NF-кB and MAPK signaling in HUVECs by activating Nrf2 signaling. There's a possibility that KNL could be developed into an anti-inflammatory drug.

Resumo Nanopartículas são consideradas opções viáveis no tratamento do câncer. Este estudo foi conduzido para investigar o efeito de nanopartículas de magnetita (MNPs) e núcleo de folato de magnetita (MFCS) em culturas de células leucêmicas e de hepatocarcinoma, bem como seu efeito no modelo animal de leucemia mielocítica aguda (LMA). Através do atual estudo, nanopartículas foram sintetizadas, caracterizadas por várias técnicas, e suas propriedades foram estudadas para confirmar sua nanoestrutura. No estudo in vivo, as nanopartículas foram avaliadas para inspecionar sua atividade citotóxica contra células SNU-182 (carcinoma hepatocelular humano), K562 (leucemia humana) e THLE2 (fígado epitelial humano normal) por meio do teste MTT. A expressão das proteínas sinalizadoras apoptóticas Bcl-2 e Caspase-3 foram inspecionadas através do método RT-PCR. Um efeito citotóxico de MNPs e MFCS foi detectado em culturas de células anteriores. Além disso, a apoptose foi identificada por meio de regulação positiva significativa da Caspase-3, com regulação negativa de Bcl-2. No estudo in vitro, a AML foi induzida em ratos por N-metil-N-nitrosoureia seguida por tratamento oral com MNPS e MFCS. Índices bioquímicos como aspartato e alanina aminotransferases e atividades de lactato desidrogenase, ácido úrico, hemograma completo e Beta-2-microglubulina foram avaliados no soro. A imunofenotipagem para detecção de CD34 e CD38 foi realizada. Fígado, rim e medula óssea foram examinados microscopicamente. A metilação do promotor Bcl-2 e os níveis de mRNA foram examinados. Embora tanto os MNPs quanto os MFCS representem uma melhora nos parâmetros bioquímicos, o MFCS os aliviou em direção ao controle normal. A atividade anticâncer de MNPs e MFCS foi aprovada especialmente para AML. Sempre, a administração de MFCS foi mais eficaz do que MNPs. O presente trabalho é um dos poucos estudos que utilizou o MFCS como agente anticâncer.

Abstract Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Abstract On March 11, 2020, the World Health Organization (WHO) declared a new coronavirus infection caused by the SARS-CoV-2 virus as a pandemic, making it the 11th pandemic of the 20th and 21st centuries. This study investigated the clinical and laboratory results (D-dimer, conventional coagulation, and HbA1c biomarker concentrations) of 150 patients (75 male and 75 female) with confirmed COVID-19 pneumonia and 50 controls (25 male and 25 female). For disease diagnosis, all COVID-19 patients were given a Real-Time Reverse Transcription Polymerase Chain Reaction Assay (RT-PCR). The findings revealed that D-dimer and HbA1c levels in COVID-19 patients were significantly higher (P 0.001) at the time of admission; In COVID-19 patients, there was also a strong correlation between D-dimer levels and HbA1c levels (P 0.001). In conclusion, COVID-19 patients are more likely to have a poor prognosis if their D-dimer and HbA1c levels remain uncontrolled over a lengthy period. To lower the likelihood of a bad prognosis in COVID-19, patients with higher levels of D-dimer and HbA1c should be continuously monitored. 11 2020 WHO (WHO SARSCoV2 SARSCoV SARS CoV 2 SARS-CoV- th st centuries Ddimer, Ddimer D dimer, dimer (D-dimer coagulation HbAc HbA c concentrations 15 (7 7 female COVID19 COVID 19 COVID-1 5 (2 female. . diagnosis RealTime Real Time RTPCR. RTPCR RT PCR (RT-PCR) P 0.001 0001 0 001 admission 0.001. conclusion period COVID19, 19, monitored 1 202 SARS-CoV ( COVID1 COVID- (RT-PCR 0.00 000 00 20 0.0 0.

Resumo Em 11 de março de 2020, a Organização Mundial da Saúde (OMS) declarou uma nova infecção por coronavírus causada pelo vírus SARS-CoV-2 como uma pandemia, tornando-a a 11ª pandemia dos séculos XX e XXI. Este estudo investigou os resultados clínicos e laboratoriais (D-dímero, coagulação convencional e concentrações de biomarcadores HbA1c) de 150 pacientes (75 homens e 75 mulheres) com pneumonia por COVID-19 confirmada e 50 controles (25 homens e 25 mulheres). Para o diagnóstico da doença, todos os pacientes com COVID-19 receberam um Ensaio de Reação em Cadeia da Polimerase com Transcrição Reversa em Tempo Real (RT-PCR). Os achados revelaram que os níveis de D-dímero e HbA1c em pacientes com COVID-19 foram significativamente maiores (P 0,001) no momento da admissão. Em pacientes com COVID-19, também houve uma forte correlação entre os níveis de D-dímero e os níveis de HbA1c (P 0,001). Em conclusão, os pacientes com COVID-19 têm maior probabilidade de ter um prognóstico ruim se seus níveis de D-dímero e HbA1c permanecerem descontrolados por um longo período. Para diminuir a probabilidade de um mau prognóstico na COVID-19, os pacientes com níveis mais altos de D-dímero e HbA1c devem ser monitorados continuamente. 1 2020 OMS (OMS SARSCoV2 SARSCoV SARS CoV 2 SARS-CoV- tornandoa tornando ª XXI Ddímero, Ddímero D dímero, dímero (D-dímero HbAc HbA c 15 (7 7 mulheres COVID19 COVID 19 COVID-1 5 (2 mulheres. . doença RTPCR. RTPCR RT PCR (RT-PCR) P 0,001 0001 0 001 admissão COVID19, 19, 0,001. conclusão período continuamente 202 SARS-CoV ( COVID1 COVID- (RT-PCR 0,00 000 00 20 0,0 0,

ABSTRACT Zinnia sp. and hairy beggartick (Bidens pilosa) plants exhibiting symptoms of possible virus infection were found in the municipality of Santa Bárbara d’Oeste, São Paulo State, Brazil. Flexuous filamentous particles and cytoplasmatic inclusions typical of potyvirus infection were observed by transmission electron microscopy, respectively, in leaf extracts and cells of symptomatic leaves. Infection of both plants with bidens mottle virus (BiMoV) was confirmed by RT-PCR using potyvirus universal primers, followed by nucleotide sequencing of the amplicons. The nearly complete genome sequence of the Brazilian isolate, named BiMoV-BR, is 9700 nucleotides long and shares 95.6 % identity with the corresponding nucleotide sequence of a BiMoV isolate from the United States. BiMoV-BR was mechanically transmitted and caused systemic infection on plants of Zinnia sp., hairy beggarstick, sunflower (Helianthus annuus), and lettuce (Lactuca sativa). Myzus persicae transmitted the virus to Zinnia sp. plants with efficacy of 8 % and 42 %, using one and ten aphids per plant, respectively. This is the first detection of BiMoV in Brazil. Further studies are necessary to evaluate the distribution of this potyvirus in the country. sp Bidens pilosa dOeste, dOeste d Oeste, Oeste d’Oeste State Brazil microscopy respectively leaves (BiMoV RTPCR RT PCR primers amplicons BiMoVBR, BiMoVBR BR, BR 970 956 95 6 95. States beggarstick Helianthus annuus, annuus , annuus) Lactuca sativa. sativa . sativa) 4 plant country 97 9

Abstract Pulmonary mesenchymal stem cells (PMSCs) are considered important in therapeutic and regenerative responses to lung injury. Despite extensive studies on the human, porcine, sheep, mouse and rat, it was found that PMSCs, which can been performed on goose, have not been well explored. Elucidation of cell differentiation, proliferation, which have not yet been well described, may promote the development of injured lung therapy. In our study, we firstly disclosed biological characteristics of spindle-shaped PMSCs from pulmonary tissues of goose embryos. Growth kinetics and counting kit-8 (CCK8) assay were employed for proliferative activity evaluation. When appropriately induced, PMSCs could differentiate into osteoblasts, adipocytes, chondrocytes, type II alveolar epithelial cells (ATII), which demonstrated that PMSCs have cross-embryonic layer differentiation potential. Besides, undifferentiated PMSCs consistently expressed MSCs associated phenotypes, such as CD166, CD90, CD44, CD29, CD71 and CD105, as identified by immunohistochemistry and RT-PCR. Karyotype analysis demonstrated that PMSCs possessed stable diploid karyotype. Collectively, we successfully established applicable methods for isolation, culture, identification and characterization of goose PMSCs which suggested a potential therapeutic use in regenerative therapy. (PMSCs injury human porcine sheep rat explored proliferation described therapy study spindleshaped spindle shaped embryos kit8 kit 8 kit- CCK8 CCK (CCK8 evaluation induced osteoblasts adipocytes chondrocytes ATII, ATII , (ATII) crossembryonic cross embryonic Besides phenotypes CD166 CD CD90 CD44 CD29 CD7 CD105 RTPCR. RTPCR RT PCR. PCR RT-PCR karyotype Collectively isolation culture (CCK (ATII CD16 CD9 CD4 CD2 CD10 CD1

Abstract Objectives To identify and understand the evidence regarding hearing changes related to acquired Dengue, Chikungunya, and Zika virus infection in adult individuals. Methods A scoping review was performed according to the recommendations of The Joanna Briggs Institute and guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews in the Embase, PubMed/Medline, ScienceDirect, Scopus, and Web of Science databases without restriction on language and year of publication. Case studies, observational studies, and clinical trials reporting hearing loss in adult subjects (>18-60 years of age) of both sexes with DENV, CHIKV, or ZIKV diagnosed by positive molecular/serological examination by RT-PCR or IgM/IgG by ELISA method were included. Results Thirteen studies met the inclusion criteria and were selected for review. The occurrence of auditory symptoms caused by arboviroses and the presence of permanent or transient sensorineural hearing loss was variable in adults. Conclusions Dengue, Chikungunya, and Zika infections in adults are associated with a variety of auditory symptoms. The frequency of permanent or transient sensorineural hearing loss is low but not negligible. Dengue Chikungunya individuals MetaAnalyses Meta Analyses Embase PubMedMedline PubMed Medline PubMed/Medline ScienceDirect Scopus publication >1860 1860 >18 60 18 (>18-6 age DENV CHIKV molecularserological molecular serological RTPCR RT PCR IgMIgG IgM IgG included negligible >186 186 >1 6 1 (>18- > (>18 (>1 (> (

Abstract Objectives Nasopharyngeal carcinoma (NPC) is an aggressive epithelial cancer. The expression of miR-186 is decreased in a variety of malignancies and can promote the invasion and metastasis of cancer cells. This study aimed to explore the role and possible mechanism of miR-186 in the metastasis and epithelial-mesenchymal transformation (EMT) of NPC. Methods The expression of miR-186 in NPC tissues and cells was detected by RT-PCR. Then, miR-186 mimic was used to transfect NPC cell lines C666-1 and CNE-2, and cell activity, invasion and migration were detected by CCK8, transwell and scratch assay, respectively. The expression of EMT-related proteins was analyzed by western blotting analysis. The binding relationship between miR-186 and target gene Zinc Finger E-Box Binding Homeobox 1 (ZEB1) was confirmed by double luciferase assay. Results The expression of miR-186 in NPC was significantly decreased, and transfection of miR-186 mimic could significantly inhibit the cell activity, invasion, and migration, and regulate the protein expressions of E-cadherin, N-cadherin and vimentin in C666-1 and CNE-2 cells. Further experiments confirmed that miR-186 could directly target ZEB1 and negatively regulate its expression. In addition, ZEB1 has been confirmed to be highly expressed in NPC, and inhibition of ZEB1 could inhibit the activity, invasion, metastasis and EMT of NPC cells. And co-transfection of miR-186 mimic and si-ZEB1 could further inhibit the proliferation and metastasis of NPC. Conclusion miR-186 may inhibit the proliferation, metastasis and EMT of NPC by targeting ZEB1, and the miR-186/ZEB1 axis plays an important role in NPC. (NPC miR186 miR 186 miR-18 epithelialmesenchymal mesenchymal (EMT RTPCR. RTPCR RT PCR. PCR RT-PCR Then C6661 C C666 C666- CNE2, CNE2 CNE 2, 2 activity CCK8 CCK assay respectively EMTrelated related analysis EBox E Box ZEB (ZEB1 Ecadherin, Ecadherin cadherin, cadherin E-cadherin Ncadherin N CNE- addition cotransfection co siZEB1 siZEB si si-ZEB miR186/ZEB1 miR186ZEB1 miRZEB 186/ZEB1 miR-186/ZEB miR18 18 miR-1 C66 (ZEB miR186/ZEB miR186ZEB 186ZEB1 186/ZEB miR1 miR- C6 186ZEB

Resumen: Brasil tiene el segundo mayor número de casos de lepra (enfermedad con carga significativa) del mundo. A pesar de los esfuerzos globales y locales para eliminar ese problema de salud pública, el diagnóstico inadecuado o tardío contribuye a perpetuar su transmisión, sobre todo entre contactos intradomiciliarios. Los exámenes como la prueba rápida de anticuerpos IgM (RT) y la reacción en cadena de la polimerasa en tiempo real (RT-PCR) se desarrollaron para superar las barreras del diagnóstico precoz de la lepra. El objetivo de este estudio fue analizar la relación de costo-efectividad de un nuevo algoritmo de diagnóstico recomendado por el gobierno brasileño para diagnosticar la lepra en contactos domiciliarios de casos confirmados de lepra, que incluye las pruebas RT y RT-PCR. Se construyó un modelo de árbol de decisión y se adoptó la perspectiva del Sistema Único de Salud (SUS) teniendo en cuenta el periodo de un año. Solo se incluyeron los costos médicos directos relacionados con los exámenes diagnósticos. Se midió la efectividad teniendo en cuenta el número de casos de lepra evitados. Se analizaron distintos escenarios. Se comparó el uso secuencial de RT, baciloscopia y RT-PCR, conforme el Ministerio de Salud recomienda, con un caso base (baciloscopia aislada), y se obtuvo un cociente de costo-efectividad incremental de USD 616,46 por cada caso de lepra evitado. El análisis de sensibilidad univariante mostró que la prevalencia de lepra entre contactos intradomiciliarios fue la variable que más influyó el modelo. Este es el primer modelo económico que analiza un algoritmo diagnóstico de lepra. Los resultados podrán ayudar los gestores a definir políticas y estrategias para erradicar la lepra en Brasil. Resumen enfermedad significativa mundo pública transmisión (RT RTPCR PCR (RT-PCR costoefectividad costo RTPCR. PCR. RT-PCR SUS (SUS año diagnósticos evitados escenarios RTPCR, PCR, recomienda aislada, aislada , aislada) 61646 616 46 616,4 evitado 6164 61 4 616, 6

Abstract: Brazil has the second largest number of leprosy cases (a disease with a significant burden) in the world. Despite global and local efforts to eliminate this public health problem, inadequate or late diagnosis contribute to perpetuate its transmission, especially among household contacts. Tests such as the rapid IgM antibody detection (RT) and real-time polymerase chain reaction (RT-PCR) were developed to overcome the challenges of early diagnosis of leprosy. This study aimed to analyze the cost-effectiveness of a new diagnostic algorithm recommended by the Brazilian government to diagnose leprosy in household contacts of confirmed leprosy cases, which includes the RT and RT-PCR tests. A decision tree model was constructed and the perspective of the Brazilian Unified National Health System (SUS) and a 1-year time horizon were adopted. Only direct medical costs related to diagnostic tests were included. Effectiveness was measured as the number of avoided undiagnosed leprosy cases. Different scenarios were analyzed. The sequential use of RT, slit-skin smear (SSS) microscopy, and RT-PCR as recommended by the Brazilian Ministry of Health was compared to a base case (isolated SSS microscopy), yielding an incremental cost-effectiveness ratio of USD 616.46 per avoided undiagnosed leprosy case. Univariate sensitivity analysis showed that the prevalence of leprosy among household contacts was the variable that influenced the model the most. This is the first economic model to analyze a diagnostic algorithm of leprosy. Results may aid managers to define policies and strategies to eradicate leprosy in Brazil. Abstract burden world problem transmission (RT realtime real RTPCR PCR (RT-PCR costeffectiveness cost effectiveness SUS (SUS 1year year 1 adopted included analyzed slitskin slit skin (SSS microscopy isolated , microscopy) 61646 616 46 616.4 most 6164 61 4 616. 6

Resumo: O Brasil tem o segundo maior número de casos de hanseníase (doença com carga significativa) do mundo. Apesar dos esforços globais e locais para eliminar esse problema de saúde pública, o diagnóstico inadequado ou tardio contribui para perpetuar sua transmissão, especialmente entre contatos intradomiciliares. Exames como o teste rápido de anticorpos IgM (RT) e a reação em cadeia da polimerase em tempo real (RT-PCR) foram desenvolvidos para superar as barreiras do diagnóstico precoce da hanseníase. Este estudo teve como objetivo analisar a relação custo-efetividade de um novo algoritmo de diagnóstico recomendado pelo governo brasileiro para diagnosticar a hanseníase em contatos domiciliares de casos confirmados de hanseníase, que inclui os testes RT e RT-PCR. Foi construído um modelo de árvore de decisão e adotada a perspectiva do Sistema Único de Saúde (SUS) considerando o período de um ano. Foram incluídos apenas os custos médicos diretos relacionados aos exames diagnósticos. A efetividade foi medida considerando o número de casos evitados de hanseníase. Diferentes cenários foram analisados. O uso sequencial de RT, baciloscopia e RT-PCR, conforme recomendado pelo Ministério da Saúde, foi comparado a um caso base (baciloscopia isolada), obtendo-se uma razão de custo-efetividade incremental de USD 616,46 por caso evitado de hanseníase. A análise de sensibilidade univariada mostrou que a prevalência de hanseníase entre contatos intradomiciliares foi a variável que mais influenciou o modelo. Este é o primeiro modelo econômico a analisar um algoritmo diagnóstico da hanseníase. Os resultados poderão auxiliar os gestores na definição de políticas e estratégias para a erradicação da hanseníase no Brasil. Resumo doença significativa mundo pública transmissão (RT RTPCR PCR (RT-PCR custoefetividade custo RTPCR. PCR. RT-PCR SUS (SUS ano diagnósticos analisados RTPCR, PCR, isolada, isolada , isolada) obtendose obtendo se 61646 616 46 616,4 6164 61 4 616, 6