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Made available in DSpace on 2022-04-29T08:32:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background & aim: Probiotics in the gut have been suggested to have a beneficial effect on anxiety response and depressive disorder. Hence we conducted a systematic review and meta-analysis to summarize the effects of probiotics associated with or without pharmacological or psychological therapies in patients with depressive and/or anxiety symptoms or disorders. Methods: We conducted searches of MEDLINE, EMBASE, CENTRAL, PsycINFO, CINAHL, ProQuest, LILACS, and Web of Science up to February 2020 to identify randomized controlled trials (RCTs) investigating the efficacy of probiotics associated with or without pharmacological or psychological therapies for patient-important outcomes including relief of depressive, anxiety and stress symptoms, cognitive functions, adverse events and quality of life. We used the GRADE approach to rate the overall certainty of the evidence by outcome. The protocol of the systematic review was registered with PROPSERO and published under the number CRD4202016329. Results: 16 RCTs including 1,125 patients proved eligible. Results suggested a significant improvement in using Beck Depression Index (MD, −3.20 [95% CI, −5.91 to −0.49], p = 0.02; I2 = 21%, p = 0.28) for depression symptoms and State-Trait Anxiety Inventory (STAI) (MD, −6.88 [95% CI, −12.35 to −1.41], p = 0.01; I2 = 24%, p = 0.25) for anxiety with overall certainty in evidence rated as moderate and low, respectively. However, Depression Scale (DASS-Depression) (MD, 2.01 [95% CI, −0.80 to 4.82], p = 0.16; I2 = 0%, p = 0.62), Montgomery-Asberg Depression Rating Scale (MADRAS) (MD, −2,41 [95% CI, −10,55 to 5,72], p = 0,56; I2 = 87%, p = 0,006), Anxiety scale (DASS-Anxiety) (MD, 0.49 [95% CI, −4.05 to 5.02], p = 0.83; I2 = 74%, p = 0.05), and Stress Scale (DASS-Stress) (MD, 0.84 [95% CI, −2.64 to 4.33], p = 0.64; I2 = 34%, p = 0.22) showed no significant decrease in the relief of depression, anxiety and stress symptoms of probiotics compared to placebo with overall certainty in evidence rated as very low for all outcomes. We also found no differences in the Beck Anxiety Index (BAI) (MD, −3.21 [95% CI, −6.50 to 0.08], p = 0.06; I2 = 0%, p = 0.88) with overall certainty in evidence rated as low. Results suggested a non-statistically significantly effect of probiotics in the adverse events outcomes. Conclusions: The current review suggests that probiotics may improve symptoms of depression and anxiety in clinical patients. However, given the limitations in the included studies, RCTs with long-term follow-up and large sample sizes are needed. Department of Biosciences and Oral Diagnosis Institute of Science and Technology Universidade Estadual Paulista McMaster Institute of Urology McMaster University St. Joseph's Healthcare Department of Community Medicine & School of Public Health PGIMER Laboratory of Experimental Research on Gynecology and Obstetrics Postgraduate Course on Tocogynecology Botucatu Medical School São Paulo State University (Unesp) Universidade Anhembi-Morumbi Campus São José Dos Campos School of Dentistry Universidade Mogi Das Cruzes High Institute of Public Health-Alexandria University Drug Information Center Tanta Chest Hospital Universidade Do Vale Do Paraíba Departament of Preventive Medicine Faculdade de Medicina - FMUSP Universidade de Sao Paulo Department of Surgical Specialties and Anesthesiology Department Botucatu Medical School UNESP - Universidade Estadual Paulista Department of Biosciences and Oral Diagnosis Institute of Science and Technology Universidade Estadual Paulista Laboratory of Experimental Research on Gynecology and Obstetrics Postgraduate Course on Tocogynecology Botucatu Medical School São Paulo State University (Unesp) Department of Surgical Specialties and Anesthesiology Department Botucatu Medical School UNESP - Universidade Estadual Paulista

Objetive. Hospital resources are insufficient given the demand for seriously ill patients with COVID-19. Proper resource management is essential to provide the best possible care. Criteria to help make appropriate and timely decisions were reviewed, following ethical principles. The prioritization of invasive ventilatory support must be done in a transparent and objective manner, comprehensively evaluating the patient and based on objective criteria such as prognostic scales, life cycle or clinical commitment. Symptomatic treatment (including palliative care) is essential in this pandemic and communication with the patient or family makes it possible to humanize the care of health personnel. Objetivo. Los recursos hospitalarios resultan insuficientes ante la demanda de pacientes graves con COVID-19. La adecuada gestión de recursos es esencial para brindar la mejor atención posible. Se revisaron criterios que ayuden a tomar decisiones adecuadas y oportunas, siguiendo principios éticos. La priorización del soporte ventilatorio invasivo debe hacerse de manera transparente y objetiva, evaluando integralmente al paciente y basado en criterios objetivos como escalas de pronóstico, ciclo de vida o compromiso clínico. El tratamiento sintomático (incluyendo cuidados paliativos) es indispensable en esta pandemia y la comunicación con el paciente o la familia permiten humanizar la atención del personal de salud.

Healthcare workers (HCWs) have been severely affected in terms of both morbidity and mortality since the beginning of the COVID-19 pandemic. During the first few months of 2021, Colombia experienced a surge in positive cases. This study aimed to evaluate the effect of vaccination on the incidence of new positive cases in HCWs.This was a retrospective cohort study of frontline employees in a network of clinics in Colombia, who were prioritized for COVID-19 vaccination from February to March 2021.Our findings were consistent with recent reports. During early 2020, the incidence of HCWs positively diagnosed with COVID-19 in Colombia was higher than that for the general population. With the start of the national vaccination program, the incidence among HCWs decreased from April 2021, while that for the general population remained relatively unchanged. Our study identified lower infection rates among HCWs during April (odds ratio [OR], 0.72 [95% CI 0.58-0.90];COVID-19 vaccination rollout in Colombia during early 2021 led to a decrease in the incidence of new positive cases among HCWs, in contrast to a continuing surge in the general population. Our findings suggested that COVID-19 vaccination provided adequate immunity, which guaranteed protection to HCWs.

La producción ovina es un renglón importante de la economía pecuaria en México, la cual puede ser mejorada con la implementación de estrategias nutricionales y biotecnológicas. Este estudio evaluó el efecto del flushing en ovejas con condición corporal media (CCM) o baja (CCB) sobre el desempeño reproductivo en un centro de apoyo a la investigación y docencia en México. Se seleccionó un grupo de 42 ovejas multíparas de razas Pelibuey, Dorper, Hampshire, East friesian y Damara asignadas a 4 tratamientos T1: (N = 10 CCM con T2: CCB¿=)(/^*ÑCA) y Baja (CCB)peci en Mexico-Ceisaflushing), T2: (N = 9 CCM sin flushing), T3: (N = 12 CCB con flushing), T4: (N = 11 CCB sin flushing). El flushing consistió en una dieta basada en sorgo (proteína cruda: 14.76), se realizaron pesajes (antes, durante y después del experimento). Se sincronizaron con un CIDR®, con retiro al día 9 y detectado el estro y monta natural inmediata. Al día 7 se aplicó 1 mL de Lutalyse® (7.5 mg PGF2α). Se evaluó la tasa de ovulación y gestación por ultrasonografía al día 45 y 75, igualmente se midió la prolificidad y fecundidad. Las medias de los pesajes fueron: antes de flushing T1: 49.08; T2: 49.15; T3: 54.46 y T4: 52.43, durante flushing T1: 48.12; T2: 47.08; T3: 55.06 y T4: 50.56 y post flushing T1: 49.42; T2: 49.11; T3: 55.00 y T4: 53.63. Las medias totales para los 3 pesajes fueron P1 = 51.28; P2 = 50.21; P3 = 51.79. En cuanto a las variables reproductivas no se encontraron diferencias estadísticamente significativas por medio de una prueba de Z; sin embargo las proporciones de los datos obtenidos muestran diferencias entre tratamientos, como los son: en la presentación del estro de los T1, T3 y T4 (90, 100 y 90.4% respectivamente) frente al T2 (55.5%); en la gestación el T1 (60%) frente a T2, T3 y T4 (44.4, 41.7 y 10% respectivamente) y respecto a la prolificidad los T1, T3 y T4 tuvieron un mayor número de crías/oveja (0.80; 1.00 y 0.90 respectivamente) respecto al T2 (0.33). El uso del flushing en animales con CCB antes de la monta o durante la sincronización con métodos farmacológicos mejora la fertilidad; pero es importante seguir supliendo los requerimientos de proteína para mejorar la fecundidad, disminuir las muertes neonatales y mejorar el peso al nacimiento de los borregos. Sheep production is an important change in the livestock economy in Mexico, the quality can be improved with the implementation of nutritional and biotechnological strategies. This study evaluated the effect of flushing in sheep with medium body condition (CCM) or Low (CCB) body condition on reproductive performance in a Research and Teaching Support Center in Mexico. A group of 42 multiparous sheep of Pelibuey, Dorper, Hampshire, East Friesian and Damara breeds assigned to 4 treatments T1: (N = 10 CCM with flushing), T2: (N = 9 CCM without flushing), T3: (N = 12 CCB with flushing), T4: (N = 11 CCB without flushing). The washing consists of a diet based on sorghum (Crude Protein: 14.76), weights (before, during and after the experiment). It is synchronized with a CIDR®, with withdrawal on day 9 and detected the estrus and immediate natural riding. On day 7, 1 mL of Lutalyse® (7.5 mg PGF2α) was applied. The rate of ovulation and pregnancy by ultrasonography at day 45 and 75 was evaluated, prolificity and fertility were also measured. The weighing averages were: before rinsing T1: 49.08; T2: 49.15; T3: 54.46 and T4: 52.43, during washing T1: 48.12; T2: 47.08; T3: 55.06 and T4: 50.56 and after discharge T1: 49.42; T2: 49.11; T3: 55.00 and T4: 53.63. The total averages for the 3 weighings were P1 = 51.28; P2 = 50.21; P3 = 51.79. Regarding reproductive variables, no statistically determined differences were found by means of a Z test; however, the proportions of the data that change the differences between treatments, such as: in the presentation of the estrus of T1, T3 and T4 (90, 100 and 90.4% respectively) versus T2 (55.5%); in pregnancy, T1 (60%) versus T2, T3 and T4 (44.4, 41.7 and 10% respectively) and regarding prolificacy, T1, T3 and T4 had a greater number of offspring/sheep (0.80, 1.00 and 0.90 respectively) with respect to T2 (0.33). The use of rinsing in animals with CCB and CCM before the mountain or during synchronization with pharmacological methods improves fertility; but it is important to continue supplying protein requirements to improve fertility, reduce neonatal deaths and improve the weight at birth of sheep.

O objetivo deste trabalho foi verificar a eficiencia da utilizacao de implantes de progesterona (P4) de quarto de uso associados ou nao a uma aplicacao de P4 injetavel. Vinte vacas da raca Nelore foram distribuidas nos grupos controle (GC): Dia (D)0 – colocacao do implante intravaginal de P4 de quarto uso, aplicacao benzoato de estradiol, D6,5 – aplicacao cloprostenol sodico, D8 – retirada implante de P4, aplicacao eCG e cipionato de estradiol; e grupo experimento (GE): o mesmo protocolo do GC com a adicao da P4 injetavel no D0. No D8 foi coletado sangue dos animais para dosagem de P4 serica. A dinâmica folicular foi acompanhada por ultrassonografia (US) na retirada do implante e a cada 12 horas ate a ovulacao, quando foi realizada a inseminacao. O diagnostico de gestacao foi realizado por US apos 40 dias da inseminacao. A media da dosagem de P4 foi de 0,63 ± 0,23 ng/mL para GC e de 2,48 ± 0,73 ng/mL para GE, resultados que diferiram significativamente. As medias do diâmetro do foliculo pre-ovulatorio (FPO) nao diferiram estatisticamente e foram de 12,52 ± 1,50 mm para GC e 12,78 ± 3,00 mm para GE. A maioria dos FPO (70%) do GC situou-se entre 11,1 e 14,4 mm de diâmetro, enquanto que no GE 50% deles apresentaram diâmetro maior que 14,4 mm. Apenas um animal do GC apresentou-se gestante. A associacao de implantes de quarto uso a progesterona injetavel foi efetiva em evitar as ovulacoes ate o momento adequado do protocolo, entretanto a quantidade excessiva de P4 circulante exerceu influencia negativa sobre as taxas de gestacao do GE.

Made available in DSpace on 2018-12-11T16:42:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The less differentiated the donor cells are used in nuclear transfer (NT), the more easily are they reprogrammed by the recipient cytoplasm. In this context, mesenchymal stem cells (MSCs) appear as an alternative to donor nuclei for NT. The amniotic fluid and adipose tissue are sources of MSCs that have not been tested for the production of cloned embryos in cattle. The objective of this study was to isolate, characterize, and use MSCs derived from amniotic fluid (MSC-AF) and adipose tissue (MSC-AT) to produce cloned calves. Isolation of MSC-AF was performed using in vivo ultrasound-guided transvaginal amniocentesis, and MSC-AT were isolated by explant culture. Cellular phenotypic and genotypic characterization by flow cytometry, immunohistochemistry, and RT-PCR were performed, as well as induction in different cell lineages. The NT was performed using MSC-AF and MSC-AT as nuclear donors. The mesenchymal markers of MSC were expressed in bovine MSC-AF and MSC-AT cultures, as evidenced by flow cytometry, immunohistochemistry, and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes, and adipocytes. Embryo production was similar between the cell types, and two calves were born. The calf from MSC-AT was born healthy, and this fact opens a new possibility of using this type of cell to produce cloned cattle by NT. University of Brasília Laboratory of Animal Reproduction Center for Technology of Zebu Dairy Breeds-CTZL Laboratory of Animal Virology and Cell Culture UNESP-University of São Paulo State Laboratory of Animal Virology and Cell Culture UNESP-University of São Paulo State

The aim of this work was to investigate the methylation and hydroxymethylation status of mesenchymal stem cells (MSC) from amniotic fluid (MSC-AF), adipose tissue (MSC-AT) and fibroblasts (FIB-control) and to verify the effect of trichostatin A (TSA) on gene expression and development of cloned bovine embryos produced using these cells. Characterization of MSC from two animals (BOV1 and BOV2) was performed by flow cytometry, immunophenotyping and analysis of cellular differentiation genes expression. The cells were used in the nuclear transfer in the absence or presence of 50 nM TSA for 20 hr in embryo culture. Expression of HDAC1, HDAC3 and KAT2A genes was measured in embryos by qRT-PCR. Methylation results showed difference between animals, with MSC from BOV2 demonstrating lower methylation rate than BOV1. Meanwhile, MSC-AF were less hydroxymethylated for both animals. MSC-AF from BOV2 produced 44.92 ± 8.88% of blastocysts when embryos were exposed to TSA and similar to embryo rate of MSC-AT also treated with TSA (37.96 ± 15.80%). However, when methylation was lower in FIB compared to MSC, as found in BOV1, the use of TSA was not sufficient to increase embryo production. MSC-AF embryos expressed less HDAC3 when treated with TSA, and expression of KAT2A was higher in embryos produced with all MSC and treated with TSA than embryos produced with FIB. The use of MSC less methylated and more hydroxymethylated in combination with embryo incubation with TSA can induce lower expression of HDAC3 and higher expression of KAT2A in the embryos and consequently improve bovine embryo production.

Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2017. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). O sucesso da clonagem por transferência nuclear (TN) está diretamente relacionado ao status epigenético da célula doadora de núcleo. Quanto menos diferenciada a célula, mais fácil é a reprogramação nuclear a um estado embrionário. Da mesma forma, o uso de drogas que atuem nos mecanismos de regulação epigenética, tal como a Tricostatina A (TSA), ajudam nessa reprogramação. O objetivo desse trabalho foi realizar a caracterização epigenética de três tipos de células doadoras de núcleo, sendo elas células-tronco mesenquimais (CTM) provenientes de duas fontes: fluido amniótico (CTM-FA) e tecido adiposo (CTM-TA), e fibroblastos (FIB) da orelha de bovinos; testar o efeito de sua utilização na TN com e sem a adição de TSA; verificar a expressão de genes ligados à epigenética: KAT2A, HDAC1 e HDAC3. Para isso, foram coletas CTM-FA por amniocentese in vivo de dois bovinos (BOV 1 e BOV 2) da raça Gir durante a gestação. Após o nascimento, foram realizadas biópsias de tecido adiposo e pele para isolamento de CTM-TA e FIB, respectivamente. As células foram cultivadas em incubadora a 38,5°C, 5% de CO2 e umidade elevada. Após isolamento, as células foram utilizadas para caracterização de sua origem mesenquimal por meio de marcadores de superfície celular verificados por citometria de fluxo e capacidade de diferenciação em adipócitos, condrócitos e osteócitos. O DNA genômico de todos os tipos celulares foi extraído pelo método de salting out e os status de metilação e hidroximetilação global do DNA foram determinados utilizando os kits MethylFlashTM Methylated DNA Quantification (Colorimetric) e MethylFlashTM Hydroxymethylated DNA Quantification (Colorimetric) (Epigentek), respectivamente. Para a clonagem, ovários foram coletados de abatedouros-frigoríficos para isolamento de ovócitos, que foram submetidos a 18 horas de maturação in vitro. Após a maturação os ovócitos foram enucleados e a célula doadora foi inserida no espaço perivitelínico. Para incorporação da célula ao citoplasma ovocitário, foi realizada a eletrofusão e então, os citoplastos foram submetidos ao cultivo com 50 nM de TSA por 20 h ou em meio sem a TSA. Após sete e oito dias de cultivo a taxa de produção de embriões foi calculada e comparada pelo teste de Tukey (p < 0,05). As culturas celulares das células provenientes do tecido adiposo e do fluido amniótico exibiram os marcadores típicos de CTM na maior porcentagem das células. Observaram-se resultados opostos entre os animais para a metilação do DNA, pois no animal BOV1, as CTM mostraram-se mais metiladas que os FIB, ao contrário do animal BOV2 onde FIB foram mais metilados que as células-tronco. Já o resultado de hidroximetilação seguiu o mesmo padrão para todas as células nos animais BOV1 e BOV2, respectivamente, sendo que CTM-FA foram as menos hidroximetiladas (0,008 e 0,002%), seguidas de FIB (0,013 e 0,009%) e CTM-TA (0,072 e 0,03%). Ao analisar as taxas de produção de embriões com sete dias de desenvolvimento, para o animal BOV1 nenhuma diferença significativa foi encontrada entre os tratamentos. Já para o animal BOV2, as CTM-FA produziram 44,92 ± 8,88% de blastocistos clonados quando os embriões foram expostos a 20 h com a TSA. Esse resultado foi superior a todos os outros tratamentos sem a TSA e semelhante ao tratamento com CTM-TA também com uso da TSA (37,96 ± 15,80%). Os embriões produzidos a partir das CTM-FA do BOV2 expressaram menos HDAC3 quando foram tratados com TSA, além disso, quando não foi utilizada TSA no cultivo, os embriões reconstruídos com FIB apresentaram menor expressão, semelhante aos embriões reconstruídos com CTM-TA. Por fim, a expressão de KTA2A foi maior em embriões tratados com TSA por 20 h quando foram utilizadas CTM, independente da fonte, como doadoras de núcleo. Estes resultados demonstram que quando as CTM foram menos metiladas que FIB, sua associação com TSA foi efetiva em aumentar a produção dos embriões bovinos clonados, entretanto, em contraste, quando as CTM foram mais metiladas sua associação com TSA gerou embriões numa proporção estatisticamente semelhante àquela obtida com CTM na ausência de TSA. Ademais, o uso da TSA foi efetivo em reduzir a expressão da HDAC3 quando CTM-FA foram utilizadas. The success of cloning by Nuclear Transfer (NT) is directly related to the epigenetic status of the nucleus donor cell. The less differentiated is the cell, the easier is nuclear reprogramming to an embryonic state. Likewise, the use of drugs acting on mechanisms of epigenetic regulation, such as Trichostatin A (TSA), would aid in reprogramming. The objective of this work was to perform the epigenetic characterization of three types of donor nucleus cells, being mesenchymal stem cells (MSC) from two sources: amniotic fluid (MSC-AF) and adipose tissue (MSC-AT), and fibroblasts (FIB) of bovine ear; test the effect of its use on NT with and without the addition of TSA; to verify the expression of genes linked to epigenetics: KAT2A, HDAC1 and HDAC3. For this, MSC-AF samples were collected by amniocentesis in vivo of two Gyr cows (BOV 1 and BOV 2) during gestation. After birth, adipose tissue and skin biopsies were collected for isolation of MSC-AT and FIB, respectively. Cells were grown in incubator at 38.5°C, 5% CO2 and elevated humidity. After isolation, the cells were used to characterize their mesenchymal origin by means of cell surface markers verified by flow cytometry and differentiation capacity in adipocytes, chondrocytes and osteocytes. The genomic DNA of all cell types was extracted by the salting out method and the methylation and hydroxymethylation status of the DNA were determined using the MethylFlash ™ Methylated DNA Quantification (Colorimetric) and MethylFlash ™ Hydroxymethylated DNA Quantification (Colorimetric) (Epigentek) kits, respectively. For cloning, ovaries were collected from slaughterhouses to isolate oocytes, which were submitted to 18 hours of in vitro maturation. After maturation, the oocytes were enucleated and the donor cell was inserted into the perivitelinic space. For incorporation of the cell into the oocyte cytoplasm, electrofusion was performed and then the cytoplasts were cultured with 50 nM TSA for 20 h or in medium without TSA. After seven and eight days of culture the rate of embryo production was calculated and compared by Tukey's test (p

Contents The objective of this work was to study cellular types that did not participated in the gastrulation process, amniotic fluid cells (AFCs) and umbilical cord cells (UCCs), in conditions of long-term culture and cryopreserved with different solutions. The AFCs and UCCs were used in a comparative study with ear fibroblast cells (EFCs) that were cultured in vitro until 20 cellular passages and cryopreserved in 10% dimethylsulphoxide (DMSO), 5% dimethyl formamide (DMF) and 7% glycerol (Gly) solutions. The cellular viability, ultrastructure, DNA fragmentation and chromosome stability were evaluated to determine the cellular type most resistant. In all cell types, it was possible to evaluate the AFCs until 15 passages and UCCs until 20 passages with different periods of cellular growth to reach the confluence phase. Solutions containing 10% DMSO ensured viability of 90.33 ± 5.58%, 90.56 ± 4.40% and 81.90 ± 3.31%, respectively for EFCs, AFCs and UCCs, being significantly more efficient and with less variation than other cryoprotectant solutions. The AFCs were more sensitive to cryopreservation and presented low viability rate at the passage 20 (17.2 ± 8.87%). There was no change in karyotype and nuclear fragmentation was low in all cellular passages studied. With the scanning electron analysis was possible the characterization of AFCs and UCCs in suspension. The three cellular types of cells presented different shapes and characteristics on the surface. The results demonstrate that bovine AFCs and UCCs can be isolated, cultured in vitro and cryopreserved in 10% DMSO, not causing damage to DNA and chromosomes. The UCCs were more resistant than AFCs in all aspects.

Wharton's jelly is a source of mesenchymal stem cells (MSCs) that had not yet been tested for bovine embryo production by nuclear transfer (NT). Thus, the objective of this study was to isolate, characterize and test MSCs derived from Wharton's jelly for embryo and pregnancy production by NT in cattle. The umbilical cord was collected during calving and cells derived from Wharton's jelly (WJCs) were isolated by explant and cultured in Dulbecco's Modified Eagle Medium. Skin Fibroblasts (FB) were isolated after 6 months of life. Morphological analysis was performed by bright field and scanning electron microscopy (SEM) during cell culture. Phenotypic and genotypic characterization by flow cytometry, immunocytochemistry, RT-PCR and differentiation induction in cell lineages were performed for WJC. In the NT procedure, oocytes at the arrested metaphase II stage were enucleated using micromanipulators, fused with WJCs or FB and later activated artificially. SEM micrographs revealed that WJCs have variable shape under culture. Mesenchymal markers of MSCs (CD29+, CD73+, CD90+ and CD105+) were expressed in bovine-derived WJC cultures, as evidenced by flow cytometry, immunocytochemistry and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes and adipocytes. After classification, the WJCs were used in NT. Blastocyst formation rate by NT with WJCs at day 7 was 25.80±0.03%, similar to blatocyst rate with NT using skin fibroblasts (19.00±0.07%). Pregnancies were obtained and showed that WJCs constitute a new cell type for use in animal cloning. RESUMO: A geleia de Wharton é uma fonte de células tronco mesenquimais (CTMs) que ainda não havia sido testada para a produção de embriões bovinos por transferência nuclear (TN). O objetivo deste estudo foi isolar, caracterizar e testar as CTMs derivadas da geleia de Wharton para produção de embriões e gestações por transferência nuclear em bovinos. O cordão umbilical foi coletado durante o nascimento e as células derivadas da geleia de Wharton (CGWs) foram isoladas por explante e cultivadas em Dulbecco's Modified Eagle Medium. Fibroblastos (FB) da pele foram isolados após 6 meses de vida. As análises morfológicas foram realizadas pelas microscopias de campo claro e eletrônica de varredura durante o cultivo celular. Caracterização fenotípica e genotípica por citometria de fluxo, imunocitoquímica, RT-PCR e indução da diferenciação em linhagens celulares foi realizada com as CGWs. No procedimento de TN, ovócitos no estágio de metáfase II foram enucleados usando micromanipuladores, fusionados com CGWs ou FB e então ativados artificialmente. Micrografias de microscopia de varredura revelaram que CGWs tiveram forma variada sob cultivo. Os marcadores mesenquimais de CTMs (CD29+, CD73+, CD90+ and CD105+) foram expressos em cultura de CGWs bovina, como evidenciado por citometria de fluxo, imunocitoquímica e RT-PCR. Quando induzidas, estas células diferenciaram-se em osteócitos, condrócitos e adipócitos. Após classificação, as CGWs foram utilizadas na TN. A taxa de formação de blastocistos por TN com CGWs no sétimo dia de cultivo foi de 25,80±0,03%, similar a produção de blastócitos por TN com fibroblastos de pele (19,00±0,07). Gestações foram obtidas e mostraram que CGWs constituem um novo tipo celular para ser usado na clonagem animal.

The design of an e-learning environment for user training in the execution of engineering projects on information technologies is presented here. Several research works claim that one of the reasons why projects fail is the lack of training of the people taking part in them. The design of the proposed e-learning environment integrates two techniques of artificial intelligence, i.e.: Intelligent tutoring systems and case-based reasoning. The results showed significant advantages when including these two techniques, specifically in the cognitive followup of the users.

Mycobacterial enoyl acyl carrier protein reductase (InhA) is a clinically validated target for the treatment of tuberculosis infections, a disease that still causes the death of at least a million people annually. A known class of potent, direct, and competitive InhA inhibitors based on a tetracyclic thiadiazole structure has been shown to have in vivo activity in murine models of tuberculosis infection. On the basis of this template, we have here explored the medicinal chemistry of truncated analogues that have only three aromatic rings. In particular, compounds 8b, 8d, 8f, 8l, and 8n show interesting features, including low nanomolar InhA IC50, submicromolar antimycobacterial potency, and improved physicochemical profiles in comparison with the tetracyclic analogues. From this series, 8d is identified as having the best balance of potency and properties, whereby the resolved 8d S-enatiomer shows encouraging in vivo efficacy.

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2013. A clonagem por transferência nuclear (TN) ainda é uma técnica com baixa eficiência em bovinos. Um dos fatores que deve ser melhorado é a fonte de células doadoras de núcleo. Quanto menos diferenciadas forem estas células, mais facilmente elas serão reprogramadas pelo citoplasma receptor, a fim de originarem embriões e bezerros a termo de uma forma saudável. Células do fluido amniótico (CFA), da geleia de Wharton do cordão umbilical (CGW) e do tecido adiposo (CTA) são fontes de células multipotentes com potencial para uma melhor reprogramação e que ainda não haviam sido testadas para a produção de embriões bovinos por meio de TN. O objetivo deste trabalho foi isolar, cultivar, criopreservar e utilizar no procedimento de TN as CFA, CGW e CTA de bovinos. Primeiramente, foram isoladas CFA de úteros de abatedouro para testar o melhor meio de cultivo (DMEM vs Amniomax Complete II), e o melhor agente crioprotetor (glicerol vs DMSO vs DMF) quanto a preservação de viabilidade celular avaliada por azul de Trypan 0,4%. Posteriormente, testou-se a possibilidade de isolamento in vivo dos tipos celulares. O cultivo foi feito por centrifugação das CFA e explante das CGW e CTA. Para comparação morfológica foi realizada a Microscopia Eletrônica de Varredura (MEV) das células em cultivo. Para avaliação de integridade de DNA realizou-se a metodologia de TUNEL. Para a análise estatística foi realizada ANOVA seguida de teste de Tukey (p

A crescente utilização das plantas para fins terapêuticos têm-se destacado em todo o mundo nas últimas décadas. Em vários países, essa forma de terapia constitui o único recurso disponível para o tratamento primário da saúde. Portanto, há necessidade de se desenvolver e comprovar a qualidade das análises químicas desses produtos. Devido à ampla utilização na medicina popular de Styrax camporum em tratamentos de transtornos gastroduodenais, os objetivos deste trabalho foram desenvolver e validar uma metodologia analítica para a detecção e a quantificação dos marcadores químicos da espécie S. camporum por CLAE-DAD, visando o controle de qualidade dessa espécie vegetal e de outras desse gênero. A cromatografia líquida de alta eficiência, no modo reverso de eluição, foi utilizada para obtenção de um cromatograma fringerprinting representativo do extrato padrão de S. camporum e para o desenvolvimento de uma condição isocrática para quantificar as nor-neolignanas presentes em espécies de Styrax. Os padrões egonol e homoegonol foram isolados de S. pohlii e usados no preparo das soluções padrão e controle de qualidade, as quais foram utilizadas para atender os requisitos: linearidade, seletividade, precisão e exatidão, limite de quantificação (LOQ) e detecção (LOD) e recuperação. Tendo em vista a comprovação da aplicabilidade do método desenvolvido, dois extratos brutos etanólicos de espécies Styrax, previamente preparados, S. pohlii e S. ferrugineus e o extrato das folhas secas de S. camporum foram utilizados para quantificar os padrões egonol (1) e homoegonol (2). A resposta do detector de UV a 311 nm foi linear de 0,36 ¿ 42,00 µg/mL para 1 e 2 e as equações da reta encontradas foram y= 123.000 x ¿ 2.743 e y= 93.490 x ¿ 10.630 (r= 0,9999 e 0,9998, CV ? 2 %), respectivamente. O método desenvolvido mostrou-se seletivo, específico, linear, preciso e exato para os dois marcadores propostos, sendo, portanto, passível de utilização no controle de qualidade de S. camporum e também de outras espécies deste gênero. The growing use of plants for therapeutic purposes has been highlighted worldwide in recent decades. In several countries, this form of therapy is the only resource available for primary health care. Therefore, there is a need to develop and prove the quality of chemical analysis of these products. Due to the widespread use in folk medicine of Styrax camporum in treatments for gastroduodenal disorders, the objectives of this work were to develop and validate an analytical methodology for the detection and quantification of chemical markers of the species S. camporum by CLAE-DAD, aiming at the control of quality of this plant species and others of that genus. High performance liquid chromatography, in the reverse elution mode, was used to obtain a fringerprinting chromatogram representative of the standard extract of S. camporum and to develop an isocratic condition to quantify the nor-neolignans present in Styrax species. The egonol and homoegonol standards were isolated from S. pohlii and used in the preparation of standard solutions and quality control, which were used to meet the requirements: linearity, selectivity, precision and accuracy, limit of quantification (LOQ) and detection (LOD ) and recovery. In order to prove the applicability of the developed method, two previously prepared crude ethanolic extracts of Styrax species, S. pohlii and S. ferrugineus and the extract of the dry leaves of S. camporum were used to quantify the egonol patterns (1) and homoegonol (2). The response of the UV detector at 311 nm was linear from 0.36 ¿42.00 µg / mL for 1 and 2 and the equations of the line found were y = 123.000 x ¿2.743 and y = 93.490 x ¿10.630 (r = 0, 9999 and 0.9998, CV - 2%), respectively. The developed method proved to be selective, specific, linear, precise and exact for the two proposed markers, being, therefore, possible to be used in the quality control of S. camporum and also of other species of this genus.

Histone deacetylase inhibitors (HDACi) have become a promising new avenue for cancer therapy, and many are currently in Phase I/II clinical trials for various tumor types. In the present study, we show that apoptosis induction and histone alterations by PCI-24781, a novel hydroxamic acid-based HDAC inhibitor, require caspase-8 and the adaptor molecule, Fas-associated death domain (FADD), in acute leukemia cells. PCI-24781 treatment also causes an increase in superoxide levels, which has been reported for other HDACi. However, an antioxidant does not reverse histone alterations caused by PCI-24781, indicating that ROS generation is likely downstream of the effects that PCI-24781 exerts on histone H3. Taken together, these results provide insight into the mechanism of apoptosis induction by PCI-24781 in leukemia by highlighting the roles of caspase-8, FADD and increased superoxide levels.