Study of ex-ovo embryonic development of Gallus gallus domesticus / Estudio inicial del desarrollo embrionario ex-ovo de Gallus gallus domesticus

SUMMARY: The domestic chicken is a species of bird that has been extensively studied in regard to its biology and as a model organism for science. The reproduction of the species is by the laying of fertilized eggs, which in a period of 21 days will develop a chick inside. Several methods have been described to develop embryos ex-ovo, allowing the observation and manipulation of the organism. This work has the propose to standardize a method that allows the development of the embryos inside the artificial incubation system, which has a low cost and is easy to make. In this work, 100 chicken eggs were used to study the effects of humidity, mineral supplementation, and the preincubation time of the egg on the incubation ex-ovo of the embryos. Embryo development was documented through the different days. Pulverized eggshell was selected as an optimal source to provide calcium, magnesium, phosphorus, and other minerals to the developing embryo. By providing 900-1200 mg of pulverized eggshell, 40 mL of the 0.001 % solution of benzalkonium chloride, and a preincubation time of approximately 56 h, the embryos were able to develop until 19 days, and even though they did not reach hatching, the incubation conditions that allowed the survival and development of embryos until late stages were achieved. Thus, due to the conditions established for calcium, humidity and preincubation time, in the present work, the chicks reached 19 days of development.

El pollo doméstico es una especie de ave que ha sido ampliamente estudiada en cuanto a su biología y como organismo modelo para la ciencia. La reproducción de la especie es por la puesta de huevos fecundados, que en un período de 21 días desarrollarán un polluelo en su interior. Se han descrito varios métodos para desarrollar embriones ex-ovo, permitiendo la observación y manipulación del organismo. Este trabajo tuvo como objetivo estandarizar un método que permita el desarrollo de los embriones dentro del sistema de incubación artificial, el cual tiene un bajo costo y es fácil de realizar. En este trabajo se utilizaron 100 huevos de gallina para estudiar los efectos de la humedad, la suplementación mineral y el tiempo de preincubación del huevo sobre la incubación ex-ovo de los embriones. El desarrollo embrionario se documentó a través de los diferentes días. Se seleccionó la cáscara de huevo pulverizada como una fuente óptima para proporcionar calcio, magnesio, fósforo y otros minerales al embrión en desarrollo. Al suministrar 900-1200 mg de cáscara de huevo pulverizada, 40 mL de la solución de cloruro de benzalconio al 0.001 % y un tiempo de preincubación de aproximadamente 56 h, los embriones lograron desarrollarse hasta los 19 días, y aunque no llegaron a eclosionar, los embriones lograron desarrollarse hasta los 19 días. Se lograron condiciones de incubación que permitieron la supervivencia y desarrollo de los embriones hasta etapas tardías. Así, debido a las condiciones establecidas de calcio, humedad y tiempo de preincubación, en el presente trabajo los pollitos alcanzaron los 19 días de desarrollo.

Optimization of callus cultures at Echinacea purpurea L. for the amount of caffeic acid derivatives

BACKGROUND: In order to produce an effective callus in Echinacea purpurea L.; determination of the explant type and growth regulators that best respond to callus induction and the optimization of the culture conditions to increase the amount of caffeic acid derivatives (CADs) in the obtained callus. CADs contents of callus cultures of E. purpurea were evaluated by establishing an effective callus induction system in vitro. RESULTS: Various medium containing different growth regulators were tested using leaf, petiole, cotyledon and root as the explants. The best callus development was achieved in MS medium with 1.0 mg l 1 2,4- D + 2.0 mg l 1 BAP in leaf, 1.0 mg l 1 NAA + 0.5 mg l 1 TDZ in petiole, 2.0 mg l 1 NAA + 1.0 mg l 1 TDZ in cotyledon and 0.5 mg l 1 NAA + 0.5 mg l 1 BAP in roots. Upon optimisation of callus growth, each type of explant was cultured for 4, 6, 8 and 10 weeks in medium for the analyses of caftaric acid, chlorogenic acid, caffeic acid and chicoric acid contents. The highest amounts of caftaric acid (4.11 mg/g) and chicoric acid (57.89 mg/g) were found from petiole explants and chlorogenic acid (8.83 mg/g) from root explants at the end of the 10-week culture time. CONCLUSIONS: As a result of the present study, the production of caffeic acid derivatives was performed by providing the optimization of E. purpurea L. callus cultures. Effective and repeatable protocols established in this study may offer help for further studies investigating the production of caffeic acid derivatives in vitro.

Presencia de bacterias en prótesis dentales durante el proceso de elaboración / Presence of bacteria in dental prostheses during the elaboration process

Existe una creciente preocupación sobre el tema de la infección cruzada en clínicas y laboratorios dentales. El laboratorio odontológico debe seguir normas de bioseguridad que garanticen a todo el equipo de salud la prevención de estas infecciones. Los técnicos que allí laboran corren el riesgo de exponer su cara a salpicaduras, así como a rocíos de sangre y saliva. Este estudio fue diseñado para saber si los laboratorios a los que recurrimos cumplen con estas normas de bioseguridad, y qué tan confiados podemos estar de la desinfección por parte de ellos, ya que las prótesis deberían estar desinfectadas correctamente antes de colocarlas en boca (AU)

There is growing concern about the issue of cross infection in dental clinics and laboratories. The dental laboratory must follow biosafety standards that guarantee the prevention of these infections to the entire health team. The technicians who work there run the risk of exposing their face to splashes and spray of blood and saliva. This study was designed to find out if the laboratories we use comply with these biosafety standards, and how confident we can be of their disinfection by them, since the prostheses should be properly disinfected before placing them in the mouth (AU)

Crop management and its effects on weed occurrence

The objective was to evaluate weed phytosociology and similarities between crop management systems in the Chapadões region. The experiment was conducted at in agricultural area located in the municipality of Chapadão do Sul, MS, during the 2016/17 harvest. Three crop managements strategies were used: (1) cotton/soybean/Urochloa, (2) millet/soybean/millet and (3) millet/soybean/crotalaria. A phytosociological survey of weeds was carried out during soybean cultivation and cover crops growth, in succession. The evaluation area for each management strategy was 0.5 ha. Soybean surveys were carried out in October and January, while the cover crop surveys were performed in February and May. The relative frequency (RF), relative density (RD), relative abundance (AR), and relative importance (RI) of weeds, Venn diagram, and Jaccard and Sorenson similarity indices were evaluated. The management area represented by the cotton/soybean/Urochloa rotation had fewer weed species than others. The species Cenchrus echinatus, Digitaria insularis, Digitaria sanguinalis, Eleusine indica and Commelina benghalensis had the highest phytosociological indeces among the monocotyledons. Attention is required for managing the dicotyledons Amaranthus deflexus, Conyza canadensis and Senna obtusifolia despite their low indices because of herbicide resistant cases. The highest indeces of similarity were found between managements areas 2 and 3, which did not rely on cotton cultivation prior to soybeans.

Eficiencia de solubilización de fósforo de aislados nativos guatemaltecos de Pseudomonas fluorescens / Phosphorus solubilization efficiency of native Guatemalan isolates of Pseudomonas fluorescens

El fósforo (P) es un elemento esencial en la producción agrícola, pero debido a su compleja dinámica en el suelo, solo una pequeña cantidad es aprovechable para las plantas, ya que la mayoría del P se encuentra en formas insolubles, especialmente, en suelos Andisoles de origen volcánico. Los microorganismos con capacidad solubilizadora de fósforo (MSF) son una alternativa para transformar el P a formas solubles y aprovechables por las plantas; además de brindar múltiples beneficios ambientales. Este trabajo identificó y evaluó in vitro, aislados nativos de Pseudomonas fluorescens Mingula, obtenidos de regiones guatemaltecas con suelos Andisoles que limitan la producción agrícola por la alta fijación de P. Se realizaron cultivos in vitro de la bacteria en medio National Botanical Research Instituteís phosphate growth (NBRIP), con fosfato tricálcico Ca3(PO4)2 como fuente de P insoluble y se midió el índice de solubilización de fósforo (ISF). Un total de 35 aislados de P. fluorescensfueron identificados y confirmados por PCR específico. El análisis de relaciones genéticas con el marcador AFLP, mostró dos grupos: el grupo A incluyó a los aislados con ISF mayores a 1.75, mientras el grupo B incluyó a aquellos con ISF menor a 1.75. La comparación de ISF entre los aislados y departamentos, demostró diferencia estadísticamente significativa (p < .001), con el aislado Pf_33 como más eficiente. Debido al potencial de solubilización de los aislados nativos del grupo genético A (ISF > 1.75), estos se recomiendan para futuras investigaciones que determinen su respuesta a condiciones de campo y estrategias para el desarrollo de biofertilizantes.

Phosphorus (P) is an essential element in agricultural production, but due to its complex dynamics in the soil, only a tiny amount is usable by plants. This is because most P is in insoluble forms, especially in volcanic Andisol soils. Microorganisms with phosphorus solubilizing capacity (MSF) are an alternative for transforming P into soluble forms usable by plants and providing multiple environmental benefits. This research identified and evaluated in vitro native isolates of Pseudomonas fluorescens Mingula, obtained from Guatemalan regions with Andisol soils that limit agricultural production due to high P fixation. In vitro cultures of the bacteria were grown on the National Botanical Research Instituteís phosphate medium (NBRIP), with tricalcium phosphate Ca3(PO4)2 as a source of insoluble P, and We measured the phosphorus solubilization index (PSI). We identified and confirmed a total of 35 isolates of P. fluorescens by specific PCR. Using the AFLP marker, genetic relationship analysis showed two groups: group A included isolates with PSI greater than 1.75, while group B included those with FSI less than 1.75. Comparing of PSI between isolates and departments showed statistically significant dif-ferences (p < 0.001), respectively, with the Pf_33 isolate as the most efficient. Because of the high solubilization potential of the native isolates of genetic group A (FSI > 1.75), We recommend future research to determine their response to field conditions and strategies for biofertilizer development.

Cultivo mediante hisopado superficial versus cultivo de la biopsia de tejidos profundos en la infección del pie diabético / Culture through superficial swab versus deep tissue biopsy in the diabetic foot infection

RESUMEN Introducción: para lograr el adecuado y precoz diagnóstico de la infección en pie diabético, es necesario la obtención de una muestra bacteriológica de calidad para la identificación del germen causal. Objetivo: identificar posibles relaciones entre los resultados obtenidos, en el cultivo realizado mediante hisopado superficial versus el obtenido mediante biopsia de los tejidos profundos en la infección del pie diabético. Materiales y métodos: se realizó un estudio explicativo observacional, longitudinal, prospectivo en el Servicio Provincial de Angiología y Cirugía Vascular del Hospital Provincial Clínico Quirúrgico Universitario "Comandante Faustino Pérez", durante un periodo de 3 años desde enero del 2016 hasta diciembre del 2018. Una selección muestral no probabilística determinó una muestra constituida por 138 extremidades en 132 pacientes con diagnóstico clínico de pie diabético infectado, que requirieron cirugía para desbridamiento de la lesión. Aceptaron ser incluidos en la investigación y para el aislamiento del germen causal fueron empleados ambos métodos de cultivo: hisopado superficial y biopsia de los tejidos profundos. Resultados: el promedio de microorganismos aislados se incrementó en relación con la severidad de la infección del pie diabético, con mayor incremento en el aislamiento hecho por el hisopado superficial. El hisopado superficial posee pobre correlación con los gérmenes aislados mediante el cultivo de la biopsia de los tejidos profundos. Conclusiones: las muestras deben ser obtenidas preferentemente por curetaje. En el diagnóstico de la infección del pie diabético es de gran utilidad, por su rapidez y concordancia con los resultados del cultivo, efectuar siempre una tinción de Gram a partir del mismo sitio (AU).

ABSTRACT Introduction: to arrive to an adequate and precocious diagnosis of the diabetic foot infection, it is necessary to obtain a qualitative bacteriological sample to identify the causing germ. Objective: to identify possible relationships between the results obtained both, in the culture made through superficial swab and the culture obtained from deep tissues biopsy in the diabetic foot infection. Materials and methods: a prospective, longitudinal, observational, explicative study was carried out in the Provincial Service of Angiology and Vascular Surgery of Provincial University Clinical Surgical Hospital "Comandante Faustino Pérez", in a period of three years, from January 2016 to December 2018. A non-probabilistic sampling choose a sample of 138 lower limbs in 132 patients with clinical diagnosis of infected diabetic foot, who required surgery for lesion debridement. They gave their consent to be included in the research; for the isolation of the casual germ were used both culture methods, superficial swab and deep tissues biopsy. Results: the average of isolated microorganism increased in relation to the severity of the diabetic food infection, with higher increase in the isolation obtained by superficial swab. The superficial swab shows poor correlation with the germ isolates by the culture the deep tissue biopsy. Conclusions: the samples should be gathered preferably by curettage. In the diagnosis of the diabetic foot infection, it is very useful, due to its speed and concordance with the culture results, to make always a Gram staining beginning from the same place (AU).

Estreptococos del grupo Streptococcus anginosus Parte I. Taxonomía, características microbiológicas e identificación / Streptococcus anginosus groupb Part I. Taxonomy, microbiological characteristics and identification / Estreptococos do grupo Streptococcus anginosus Parte I. Taxonomia, características microbiológicas e identificação

Resumen Los estreptococos del grupo Streptococcus anginosus (EGA), también llamados "Streptococcus milleri" fueron reconocidos como parte de los estreptococos del grupo viridans (EGV) desde principios del siglo XX. Sin embargo, su rol como patógenos humanos comenzó a destacarse recién en la década de 1970. Esta actualización consta de tres partes: en esta primera parte se tratarán los aspectos taxonómicos y microbiológicos así como los métodos de identificación de los EGA. El crecimiento de estas bacterias es relativamente lento, las colonias son pequeñas, incluso a las 48-72 horas de incubación y la mayoría de las cepas despide un olor a caramelo característico cuando crecen en agar sangre. Su crecimiento es estimulado en una atmósfera con 5% de CO2. Últimamente, con el reconocimiento de la asociación de los EGA con episodios indeseables en pacientes con fibrosis quística se han desarrollado medios selectivos para poner de manifiesto su presencia en las vías aéreas. Los métodos fenotípicos e incluso algunos genotípicos carecen de precisión para identificar las tres especies del grupo (Streptococcus anginosus, Streptococcus constellatus y Streptococcus intermedius) e incluso pueden fallar en su clasificación a nivel de grupo. Dentro de los métodos moleculares, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) no puede ser tomado como referencia para llegar a subespecie, pero sí es muy eficiente en la identificación a nivel de especie. Para algunos autores la secuenciación del gen sodA podría ser una buena opción, pero el gold standard es el multilocus sequence analysis (MLSA).

Abstract Streptococci from the Streptococcus anginosus group (SAG), also called "Streptococcus milleri", have been recognized as belonging to the viridans group (VGS) since the beginning of the 20th century. Their role as human pathogens, however, only began to emerge in the 1970s. This review consists of three parts: the first part will deal with the taxonomic and microbiological aspects and the identification methods of SAGs. The growth of these bacteria is relatively slow; the colonies are small even after 48-72 hours of incubation and most of the strains give off a characteristic caramel odor when they grow on blood agar. Their growth is stimulated in an atmosphere with 5% CO2. Lately, with the recognition of the association of SAGs with undesirable episodes in patients with cystic fibrosis, selective media have been developed to reveal their presence in the airways. Phenotypic and even some genotypic methods lack precision in identifying the three species in the group (Streptococcus anginosus, Streptococcus constellatus, and Streptococcus intermedius) and may even fail to classify at the group level. Among the molecular methods, MALDI-TOF MS cannot be taken as a reference to arrive at subspecies, but it is very efficient to identify at the species level. For some authors, sequencing the sodA gene may be a good option, but the gold standard is multilocus sequence analysis (MLSA).

Resumo Os estreptococos do grupo Streptococcus anginosus (EGA), também chamados de "Streptococcus milleri", foram reconhecidos como pertencentes ao grupo viridans (EGV) desde o início do século XX. Seu papel como patógenos humanos, no entanto, só começou a surgir na década de 1970. Esta atualização consiste em três partes: nesta primeira parte, trataremos dos aspectos taxonômicos e microbiológicos e dos métodos de identificação dos EGAs. O crescimento dessas bactérias é relativamente lento, as colônias são pequenas mesmo após 48-72 horas de incubação e a maioria das cepas emitem um cheiro de caramelo característico quando crescem em ágar sangue. Seu crescimento é estimulado em uma atmosfera com 5% de CO2. Ultimamente, com o reconhecimento da associação dos EGAs com episódios indesejáveis em pacientes com fibrose cística, foram desenvolvidos meios seletivos para revelar sua presença nas vias aéreas. Os métodos fenotípicos e mesmo alguns genotípicos carecem de precisão na identificação das três espécies do grupo (Streptococcus anginosus, Streptococcus constellatus e Streptococcus intermedius) e podem até falhar em sua classificação em nível de grupo. Entre os métodos moleculares, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) não pode ser tomado como referência para chegar a subespécie, mas é muito eficiente na identificação em nível de spécie. Para alguns autores, o sequenciamento do gene sodA poderia ser uma boa opção, mas o padrão-ouro é a análise de sequência multilocus (MLSA).

Diagnóstico de la Tricomonas vaginalis en la mujer / Diagnosis of Trichomonas vaginalis in women

OBJETIVO: revisar los diferentes métodos de diagnóstico de la tricomoniasis vaginal disponibles hasta el presente. MATERIALES Y MÉTODOS: se revisó la bibliografía latinoamericano e internacional a través de los sitios electrónicos de Pub-Med y Scielo. RESULTADOS: la Tricomonas vaginalis es considera como la enfermedad de transmisión sexual no viral, curable más frecuente y prevalente en el mundo. Se revisan los diferentes de métodos para diagnosticar la presencia de la tricomonas vaginalis en pacientes femeninos con síntomas y signos de la infección producida por el protozoario flagelado. CONCLUSIONES: se revisaron los diferentes métodos de diagnostico de la infección producida por la Tricomonas vaginalis en pacientes femeninas, desde los clásicos hasta los más actuales que emplean alta tecnología.

OBJECTIVE: to review the different diagnostic methods of Trichomonas vaginal available at the present time. MATERIAL AND METHOD: it was reviewed the Latin-American and international bibliography using the Pub-Med and Scielo web sites. RESULTS: Trichomonas vaginalis is considered the most common and prevalent sexual transmitted disease curable and non-viral worldwide. It was reviewed the different methods to diagnose the presence of Trichomonas vaginalis in female patients with symptoms and signs of infection produces by the flagellate protozoa. CONCLUSION: Different methods of diagnosis of the infection produced by Trichomonas vaginalis, since the classics to the most current methods that use high technology, were reviewed.

Development of Ctenocephalides felis felis (Siphonaptera: Pulicidae) in different substrates for maintenance under laboratory conditions / Desenvolvimento de Ctenocephalides felis felis (Siphonaptera: Pulicidae) em diferentes substratos para manutenção em condições laboratoriais

Abstract The aim of this study was to evaluate the efficiency of different substrates for larval development of Ctenocephalides felis felis during its biological cycle. Eight hundred eggs of C. felis felis from a flea maintenance colony were used. Different diets were formulated, in which the main substrates were meat flour, powdered milk, sugar, lyophilized bovine blood, tick metabolites and lyophilized egg. The flea eggs were placed in test tubes (10 per tube) and approximately 2 g of the diet to be tested was added to each tube. There were 10 replicates for each substrate. After 28 days, each tube was evaluated individually for the presence of pupae and emerged adults. The following percentages of the larvae completed the cycle to the adult stage: 67% in diets containing tick metabolites; 55%, meat flour; 39%, dehydrated bovine blood; 14%, powdered milk; and less than 1% in diets containing sugar, lyophilized bovine blood, lyophilized egg or wheat bran. It was concluded that among the diets tested, the one constituted by tick metabolites as the substrate was shown to be the most satisfactory for maintaining a laboratory colony of C. felis felis, followed by the one containing meat flour.

Resumo Este trabalho teve como objetivo avaliar a eficiência de diferentes substratos no desenvolvimento larval de Ctenocephalides felis felis durante seu ciclo biológico. Foram utilizados 800 ovos de C. felis felis, oriundos de colônia de manutenção de pulgas. Diferentes dietas foram formuladas, contendo como substratos principais a farinha de carne, leite em pó, açúcar, sangue bovino liofilizado, metabólitos de carrapato e ovo liofilizado. Foram distribuídos 10 ovos por tubo de ensaio, aos quais foram acrescidos as dietas a serem testadas, realizando-se10 repetições para cada substrato. Após 28 dias, cada tubo foi avaliado individualmente pela presença de pupas e adultos emergidos. Nas dietas que continham metabólitos de carrapato, 67% das larvas completaram o ciclo até a fase adulta; 55% nas que continham farinha de carne; 39% contendo sangue bovino desidratado; 14% com leite em pó, e menos de 1% em dietas contendo açúcar, sangue bovino liofilizado, ovo liofilizado e farelo de trigo. Conclui-se que, entre as dietas testadas, a constituída por metabólitos de carrapato como substrato, mostrou-se a mais satisfatória para a manutenção de colônia laboratorial de C.felis felis, seguida da que continha farinha de carne.

Phytochemical Screening of Callus and Cell Suspensions Cultures of Thevetia peruviana

Abstract Thevetia peruviana is an ornamental shrub grown-up in many tropical region of the world. This plant produces secondary metabolites with biological properties of interest for the pharmaceutical industry. The objective was to determine the secondary metabolites profile of callus and cell suspension cultures of T. peruviana and compare them with those from explant (fruit pulp). Extracts in 50% aqueous ethanol and ethyl acetate were prepared. The phytochemical analysis was performed using standard chemical tests and thin layer chromatography. In addition, total phenolic and flavonoids compounds (TPC and TFC), total cardiac glycosides (TCG) and total antioxidant activity (TAA) was determined during the cell suspension growth. Phenolic chemical profile was also analyzed by high performance liquid chromatography (HPLC). Common metabolites (alkaloids, amino acids, antioxidants, cardiac glycosides, leucoanthocyanidins, flavonoids, phenols, sugars and triterpenes) were detected in all samples. The maximum production of extracellular TCG, TPC, TFC and TAA in cells suspensions were at 6-12 days; in contrast, intracellular content was relatively constant during the exponential grown phase (0 to 12-days). HPLC analysis detected one compound with retention time at 11.6 min; this compound was tentatively identified as dihydroquercetin, a flavonoid with anti-cancer properties. These results provide evidence on the utility of the in vitro cell cultures of T. peruviana for valuable pharmaceutical compounds production.

Megacariopoyesis humana in vitro: determinación de la concentración óptima de trombopoyetina / In vitro human megacaryocytopoiesis: determination of thrombopoietin optimal concentration / Megacariópoiesis humana in vitro: determinação da concentração ótima da trombopoietina

El estudio de la megacariopoyesis humana se ha visto obstaculizado por la relativa escasez de megacariocitos en la médula ósea (0,05-0,2 % de las células medulares), lo que ha llevado a la optimización de protocolos de expansión in vitro a partir de precursores de diversos orígenes (cordón umbilical, médula ósea y sangre periférica con o sin movilización previa). Los cultivos celulares a partir de precursores han permitido la producción y el estudio tanto de megacariocitos así como de proplaquetas y plaquetas Sin embargo, la producción in vitro óptima de megacariocitos que culminen todos los estadios de diferenciación es un reto aún no resuelto. En este trabajo reportamos los hallazgos concernientes a la determinación de las condiciones y concentraciones de trombopoyetina para lograr una óptima relación entre la cantidad de trombopoyetina empleada y el porcentaje y grado de diferenciación megacariocítica en muestras obtenidas de cinco donantes alogénicos aceptados para trasplante de médula ósea.

The study of human megakaryocytopoiesis has been hampered by the relative scarcity of megakaryocytes in bone marrow (0.05-0.2 % of medullary cells), which has led to the optimization of protocols of in vitro expansion of precursors from diverse sources (umbilical cord, bone marrow and peripheral blood with or without previous mobilization). Cell cultures from different precursors have allowed the production and study of megakaryocytes as well as proplatelets and platelets. However, the in vitro production of megakaryocytes that culminate all stages of differentiation is a challenge that has not yet been resolved. In this work we report the findings related to the determination of thrombopoietin treatment conditions and concentrations to achieve an optimal relationship between the amount of thrombopoietin and the percentage and degree of megakaryocytic differentiation in five allogeneic donors that were accepted for bone marrow transplantation.

O estudo da megacariopoiese humana tem sido dificultado pela relativa escassez de megacariócitos na medula óssea (0,05-0,2 % das células medulares), o que levou à otimização dos protocolos de expansão in vitro a partir de precursores de diversas origens (cordão umbilical, medula óssea e sangue periférico com ou sem mobilização prévia). Culturas de células a partir de precursores permitiram a produção e o estudo tanto de megacariócitos e de proplaquetas e plaquetas. No entanto, a produção ótima in vitro de megacariócitos que culminam em todas as fases de diferenciação é um desafio ainda não resolvido. Neste trabalho, relatamos as descobertas relativas à determinação das condições e concentrações de trombopoietina para obter uma relação ótima entre a quantidade de trombopoietina usada e a taxa e o grau de diferenciação megacariocítica em amostras obtidas de cinco doadores alogênicos aceitos para transplante de medula óssea.

Blefarite: epidemiologia, etiologia, apresentações clínicas, tratamento e evolução de nossos pacientes / Blepharitis: epidemiology, etiology, clinical presentations, treatment and evolution of our patients

Resumo Objetivo: A blefarite é uma das condições mais comumente encontradas na prática oftalmológica e se constitui em uma causa frequente de irritação e desconforto ocular. Por ser uma doença de difícil tratamento, os autores buscaram compreender melhor a epidemiologia, etiologia, apresentações clínicas, tratamento e evolução de seus pacientes, visando maior sucesso terapêutico. Métodos: Foram avaliados retrospectivamente e transversalmente o prontuário de 124 pacientes do Centro de Oftalmologia Tadeu Cvintal, os quais apresentavam blefarite e foram submetidos à classificação de gravidade e coleta de secreções palpebrais para cultura bacteriana e antibiograma. Resultados: A media da idade dos pacientes foi de 67,4 anos, o sexo feminino foi responsável por 70 (56,4%) casos e o masculino por 54 (43,5%). Quanto à gravidade da doença, constatou-se 71 casos de blefarite leve (56,8%), 52 (41,6%) com intensidade moderada e 2 (1,6%) casos graves. Avaliando o seguimento do tratamento da doença, foi observado que 103 (82,4%) pacientes não retornaram para avaliar o resultado do tratamento e apenas 22 (17,6%) retornaram. Em relação às culturas realizadas, 82 (66,1%) não apresentaram crescimento microbiano. Dentre as 42 (33,8%) amostras positivas, os Staphylococcus coagulase negativo foram os mais prevalentes, sobretudo os Staphylococcus epidermidis, responsável por 35 (83,3%) delas. Quanto à sensibilidade aos antibióticos, os agentes de nossa amostra demonstraram maior resistência à Penicilina, Eritromicina e Ciprofloxacino e 100% de sensibilidade à Linezolida, Vancomicina e Daptomicina. Conclusão: Conhecendo melhor as características epidemiológicas da blefarite e a sensibilidade antimicrobiana das bactérias envolvidas, é possível oferecer tratamentos mais eficazes.

Abstract Objective: Blepharitis is one of the most commonly encountered conditions in ophthalmic practice and is a frequent cause of eye irritation and discomfort. Being a difficult to treat disease, the authors sought to better understand the epidemiology, etiology, clinical presentations, treatment and evolution of their patients, aiming at greater therapeutic success. Methods: The medical records of 124 patients of Centro de Oftalmologia Tadeu Cvintal who had blepharitis were retrospectively and cross-sectionally evaluated and underwent severity classification and collection of eyelid secretions for bacterial culture and antibiogram. Results: The mean age of the patients was 67.4 years, females accounted for 70 (56.4%) cases and males for 54 (43.5%). Regarding the severity of the disease, there were 71 cases of mild blepharitis (56.8%), 52 (41.6%) with moderate intensity and 2 (1.6%) severe cases. Evaluating the follow-up of treatment of the disease, it was observed that 103 (82.4%) patients did not return to evaluate the treatment outcome and only 22 (17.6%) returned. In respect of the cultures performed, 82 (66.1%) did not show microbial growth. Among the 42 (33.8%) positive samples, coagulase-negative staphylococci were the most prevalent, especially Staphylococcus epidermidis, responsible for 35 (83.3%) of them. As for antibiotic sensitivity, the agents in our sample showed greater resistance to Penicillin, Erythromycin and Ciprofloxacin and 100% sensitivity to Linezolid, Vancomycin and Daptomycin. Conclusion: By better understanding the epidemiological characteristics of blepharitis and the antimicrobial sensitivity of the bacteria involved, it is possible to offer more effective treatments.

Micropropagacion in vitro de Rosa rosa sp. a partir de yemas axilares y respuesta callogénica / Rose rosa sp. in vitro micropropagation from axillary buds and callogenic response

El objetivo de este trabajo fue obtener plántulas de Rosa sp y respuesta callogénica a partir de yemas axilares y hojas. Se utilizó Murashige y Skoog (MS) al 100%, como medio de cultivo para la fase de establecimiento, adicionado con myo-inositol 0,1 mg/L, ácido ascórbico 0,1 mg/L, tiamina 0,2 mg/L, sacarosa 30 g, benzylaminopurina (BAP) 0,4 mg/L, kinetina 0,3 mg/L, ácido naftaleno-acético (ANA) 0,3 mg/L, y Agar sigma 6 g; ajustando el pH a 5,8; esterilizados en autoclave a 120°C bajo 15 lb de presión. El análisis de varianza para la fase de multiplicación y enraizamiento determinó que existen diferencias altamente significativas para la variable altura ( p=0,000), para determinar en cuáles medias son significativamente diferentes se realizó la prueba de Kruskal ­ Wallis. Para las variables presencia de raíz (p = 0,3398), número de hojas (p = 0,4775) y formación de callos (p = 0,1964), sin diferencias estadísticamente significativas.

The objective of this work was to obtain Rosa sp seedlings and callogenic response from axillary buds and leaves. Murashige and Skoog (MS) 100%, was used as culture medium for the establishment phase, added with myo-inositol 0.1 mg / L, ascorbic acid 0.1 mg / L, thiamin 0.2 mg / L, sucrose 30 g, benzylaminopurine (BAP) 0.4 mg / L, kinetin 0.3 mg / L, indole-acetic acid (IAA) 0.3 mg / L, and Sigma agar 6 g; adjusting the pH to 5.8; sterilized in an autoclave at 120 °C under 15 lb of pressure. The analysis of variance for the phase of multiplication and rooting determined that there are highly significant differences for the variable height (p = 0.000), in order to determine which means are significantly different, the Kruskal - Wallis test was performed. For the variables presence of root (p = 0. 3398), number of leaves (p = 0.4775) and callus formation (p = 0.1964), without statistically significant differences.

Applications of photobiomodulation in hearing research: from bench to clinic

Hearing loss is very common and economically burdensome. No accepted therapeutic modality for sensorineural hearing loss is yet available; most clinicians emphasize rehabilitation, placing hearing aids and cochlear implants. Photobiomodulation (PBM) employs light energy to enhance or modulate the activities of specific organs, and is a popular non-invasive therapy used to treat skin lesions and neurodegenerative disorders. Efforts to use PBM to improve hearing have been ongoing for several decades. Initial in vitro studies using cell lines and ex vivo culture techniques have now been supplanted by in vivo studies in animals; PBM protects the sensory epithelium and triggers neural regeneration. Many reports have used PBM to treat tinnitus. In this brief review, we introduce PBM applications in hearing research, helpful protocols, and relevant background literature.

Stem cell-based therapy for inflammatory bowel disease

Inflammatory bowel disease (IBD) is an idiopathic, multi-etiological disease characterized by inflammation and mucosal destruction of the gastrointestinal tract. Despite the remarkable advance in immunomodulating therapies, there still remains a certain population of patients who are refractory to conventional as well as biologic therapies and fail to achieve mucosal healing. To improve the prognosis of those patients, at least 2 types of stem cells have been tested for their potential therapeutic use. Transplantation of hematopoietic stem cells or mesenchymal stem cells have been tested in several clinical studies, but their beneficial effect still remains controversial. In this review, we would like to overview the recent clinical challenges of stem cell-based therapies in IBD and also introduce our new therapeutic plan of intestinal stem cell transplantation for IBD, based on our ex vivo intestinal organoid culture technique.

Assessment of different methods for the detection of biofilm production in coagulase-negative staphylococci isolated from blood cultures of newborns

Abstract INTRODUCTION: Coagulase-negative staphylococci (CoNS) are a frequent cause of bacteremia, especially in neonates. The major virulence determinant in CoNS is the ability to produce biofilms, which is conferred by the icaADBC genes. This study aimed to assess different methods for the detection of biofilm formation in 176 CoNS isolates from blood cultures of newborns. METHODS: The presence of the icaACD genes was assessed by polymerase chain reaction (PCR), and biofilm formation was assessed on congo red agar (CRA), by the tube method (TM), and on tissue culture plates (TCP). RESULTS: Of the 176 CoNS isolates, 30.1% expressed icaACD and 11.4% expressed icaAD. The CRA assay and TM showed that 42% and 38.6% of the isolates were biofilm producing, respectively. On TCP, 40.9% of the isolates produced biofilms; 21% were weakly adherent and 19.9% were strongly adherent. When compared to the gold standard technique (PCR), the CRAassay showed 79% sensitivity and 84% specificity (kappa = 0.64), TM showed 78% sensitivity and 89% specificity (kappa = 0.68), and TCP showed 99% sensitivity and 100% specificity (kappa = 0.99). CONCLUSIONS: In this study, ~42% of CoNS isolates produced biofilms, and the presence of icaACD was associated with a greater capacity to form biofilms. Compared to the other phenotypic methodologies, TCP is an ideal procedure for routine laboratory use.

Culture and patients' perception to maxillofacial surgical practices in kano, Nigeria

Objective: Culture is the way of life of a people; and is an integral component of their day-to-day existence. It influences the daily routine of a people, including their diet, dressing, religious disposition, and surprisingly, the degree to which orthodox medical practices impact their daily lives. Appreciating underlying cultural context will help health care workers influence patient's perceptions, especially where cultural practices are not in tandem with medical best practices. This is important, for example, in administration of informed consent for surgery. This study explored cultural beliefs of patients in relation to some common maxillofacial practices in Kano, Nigeria. Methods: Patient's perceptions on oral cancers, use of nasogastric tubes, and tooth extraction was conducted amongst patients attending maxillofacial outpatient clinic of a tertiary Nigerian hospital from January to December 2015 using a non-structured, interviewer-administered questionnaire. Results: Seventy-seven (77) patients were surveyed (52 males and 25 females), with ages ranging from 16 to 75 years. Most were aged 51-60 years (44.2%). Only 6.5% of respondents had higher than secondary education. Responses to the aetiology and treatment of oral cancers, use of nasogastric tubes for maxillofacial surgery patients and extraction of teeth showed cultural perceptions usually at variance with medical best practices. Conclusion: Patients' expectations and fears of maxillofacial surgery procedures are affected by their cultural beliefs. Proper acceptance of this, combined with targeted education and counselling may enhance patient's co-operation and acceptance of necessary surgical procedures when orthodox medical care is sought

Bacterial infection monitoring in the early period after liver transplantation

PURPOSE: Infection remains the main cause of morbidity and mortality in liver transplantation (LT) recipients; however infection is notoriously difficult to diagnose because its usual signs and symptoms of infection may be masked or absent. This study comprises an analysis of bacterial infections in the early period after LT. METHODS: This is a study of 129 adults who underwent LT from January 2013 to December 2013, and it includes patients who were followed daily from the day of transplantation to 1-week posttransplantation using bacteriological cultures of blood, urine, sputum, and drained ascites. RESULTS: The following factors were significantly different between the positive and negative culture groups: living donor LT vs. deceased donor LT (odds ratio [OR], 3.269; P = 0.003), model for end-stage liver disease score (OR, 4.364; P < 0.001), and Child-Pugh classification (P = 0.007). Neither positive culture nor negative culture was associated with infection within 4 weeks of surgery (P = 0.03), and most events were due to surgical complications (75%). CONCLUSION: Since the full effect of immunosuppression is not yet present during the first month after LT, we suggest that the number of bacterial culture test could be reduced such that they are performed every other day depending on patient's situation.

Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis

ABSTRACT Early diagnosis of tuberculosis is of major clinical importance. Among 4733 clinical specimens collected from 3363 patients and subjected to Ziehl-Neelsen microscopy, 4109 were inoculated onto Löwenstein-Jensen slants and 3139 in Bactec/9000MB. Polymerase Chain Reaction (PCR) was performed in 3139 specimens, whereas, a genotypic assay was directly applied in 93 Mycobacterium tuberculosis complex PCR-positive for isoniazid and rifampicin resistance detection specimens (GenoType MTBDRplus). Recovered M. tuberculosis isolates (64) as well as, 21 more sent from Regional Hospitals were tested for antimycobacterial resistance with a phenotypic (manual MGIT-SIRE) and a genotypic assay (GenoType MTBDRplus). PCR in the clinical specimens showed excellent specificity (97.4%) and accuracy (96.8%), good sensitivity (70.4%), but low positive predictive value (40.3%). MGIT-SIRE performed to M. tuberculosis did not confer a reliable result in 16 isolates. Of the remaining 69 isolates, 15 were resistant to streptomycin, seven to isoniazid, seven to ethambutol and five to rifampicin. GenoType MTBDRplus correctly detected isoniazid (seven) and rifampicin-resistant M. tuberculosis strains (five), showing an excellent performance overall (100%). Susceptibility results by the molecular assay applied directly to clinical specimens were identical to those obtained from recovered isolates of the corresponding patients. Combining molecular and conventional methods greatly contribute to early diagnosis and accurate susceptibility testing of tuberculosis.