Producción de plásmidos recombinantes para su uso como controles de la técnica RT-PCR para el diagnósticode los virus Chikungunya y Zika / Production of recombinant plasmids as controls of diagnosis technique (RTPCR) of Chikungunya and Zika viruses

El diagnóstico molecular de arbovirus es indispensable para identificar agentes etiológicos, particularmente en zonas endémicas para al menos uno de ellos. Estas deben ser validadas con controles positivos, los cuales están clásicamente representados por virus vivos, cuya obtención puede ser riesgosa, laboriosa y costosa. El objetivo de este estudio fue producir plásmidos recombinantes para su uso como controles positivos en la validación de la técnica RT-PCR para el diagnóstico de los virus Chikungunya (CHIKV) y Zika (ZIKV). A partir de los ARN extraídos de los virus [CHIKV (LARD809-GC) y ZIKV (MR766)] se obtuvieron por RT-PCR fragmentos parciales de ADN correspondientes a secuencias nucleotídicas de los genes E1 y NS5 de los virus Chikungunya y Zika, respectivamente, para serclonados en el plásmido comercial pGEM®-T Easy. La clonación se confirmó mediante PCR de colonias y PCR de ADN plasmídicos extraídos a partir de las colonias recombinantes. Se logró la producción de dos plásmidos recombinantes CHIKV-E1/pGEM®-T Easy y ZIKV-NS5192/pGEM®-T Easy con cada una de las secuencias especificadas, para su uso en la validación y control de las técnicas moleculares descritas en este reporte, para el diagnóstico de agentes virales CHIKV y ZIKV, evitando la manipulación de cultivos celulares y garantizando una fuente confiable de controles positivos(AU)

The use of molecular techniques for the viral diagnosis requires the use of positive controls.Classically, the controls are live viruses, whose manipulation may be risky, laborious and expensive. The objective of this study was produced recombinant plasmids to obtain cloned sequences of Chikungunya (CHIKV) and Zika (ZIKV) virus for their use as controls in the specificdiagnostic by RT-PCR. DNA fragments were obtained fromRNA [CHIKV (LARD809-GC) and ZIKV (MR766)] using specific primers to amplify the nucleotide sequences from fragments of Envelope 1 protein (E1) of CHIKV and Non Structural 5 protein (NS5) of ZIKV genomes. The 548 bp (CHIKV) and 192 bp(ZIKV) bands were purified from agarose gel and ligations were performed with the cloning vector pGEM®-T Easy. The Escherichia coli XL1-Blue MRF` cells were transformed with the ligation mixture, the recombinant colonies were identified by colony PCR using the specific primers to the specific viral agent. One recombinant colony from CHIKV and six recombinant colonies from ZIKV were obtained from which plasmidic DNAs was extracted. The plasmidic DNAs were used as reaction controls in CHIKV and ZIKV RT-PCR, obtaining the characteristic bands. The cloning of the sequences was successful to produce the recombinant plasmids (CHIKV-E1/pGEM®-T Easy y ZIKV-NS5192/pGEM®-T Easy) to use in the validation of RT-PCR techniques(AU)

Treinamento muscular inspiratório e atividades de vida diária em idosas com doença de Parkinson / Inspiratory muscle training and activities of daily living of elderly with Parkinson disease

Introdução: Parkinson é uma condição degenerativa caracterizada por sintomas como bradicinesia, tremor, rigidez, diminuição da força muscular e alterações cognitivas. Objetivo: Avaliar a força muscular inspiratória máxima, o efeito do treinamento muscular inspiratório pré e pós-treinamento muscular, a presença de dispneia, o padrão ventilatório e a independência funcional em idosas institucionalizadas com síndrome parkinsoniana. Material e métodos: Uma série de casos, com oito idosas, em que se avaliou presença de dispneia, padrão ventilatório, índice de Katz no pré-treinamento e manovacuometria antes e depois do fortalecimento da musculatura inspiratória. Utilizou-se Threshold® IMT, 3x na semana por 30 minutos em dois meses. A carga inspiratória iniciou com 30% da PImáx com incremento de 10% a cada nove dias chegando a 50 %. Os dados foram analisados com estatística descritiva e apresentados em média e desvio padrão e o Teste t para avaliar a diferença estatisticamente significativa. Resultados: Obteve-se melhora da PImáx com diferença estatisticamente significativa (p = 0,004), porém não alcançaram o predito para a idade das mesmas, no Índice de Katz 50% das idosas eram parcialmente independentes, 37,5% independentes e 12,5% totalmente dependente. Conclusão: As voluntárias apresentaram algum grau de dependência em suas atividades de vida diária, e apresentaram ganho na PImáx após o treinamento. (AU)

Introduction: Parkinson's is a degenerative condition characterized by symptoms as bradykinesia, tremor, rigidity, decrease of muscular force and cognitive alterations. Objective: To evaluate the maximum inspiratory muscle strength, the effect of inspiratory muscle training, before and after training muscle, the presence of dyspnea, the ventilatory standard and the functional independence in institutionalized elderly woman with parkinsonian syndrome. Methods: A series of cases, with eight elderly women, which the presence of dyspnea, respiratory pattern, Katz index, pre training and manovacuometry were evaluated before and after strengthening inspiratory muscles. It was used Threshold ® IMT, 3x a week for 30 minutes in two months. The inspiratory load started with 30% of PImáx with an increment of 10% every nine days reaching 50%. Data were analyzed with descriptive statistics and presented as mean and standard deviation and t test to assess the difference statistically significant. Results: We observed that Pimáx improvement was statistically significant (p = 0.004), but did not reach the predicted for the age of the same, Katz Index of 50% of the elderly women are partly independent, 37.5% independent and 12.5% totally dependent. Conclusion: The volunteers showed some degree of dependence in activities of daily living, and gain in PImáx after training.(AU)

Valoración terapéutica del agua de mar en modelos experimentales como terapia complementaria en anemia / Valuation therapeutic seawater in experimental models as complementary therapy in anaemia

Introducción: la anemia es un problema de salud pública a nivel mundial, según la Organización Mundial de la Salud afecta en todo el mundo a 1620 millones de personas. En América Latina y el Caribe el 31% de la población presenta problemas de anemia asociadas a dietas deficientes de hierro y relacionadas con malnutrición. Objetivo: evaluar los efectos terapéuticos del agua de mar isotónica microfiltrada al frío como terapia complementaria en modelos biológicos con anemia inducida experimentalmente por dietas pobres en proteínas, vitaminas y minerales, y por medicamentos. Materiales y métodos: se contó durante ocho semanas con 50 ratones cepas NMRI, de ambos géneros y en edad adulto-joven. Se dividieron en cuatro grupos experimentales de 10 ratones cada uno, incluyendo un grupo control. Fueron evaluados al inicio y final del experimento los parámetros hematológicos, pH salival, cobre y zinc. Al final del experimento fueron sacrificados por dislocación cervical y autopsiados para examinar bazo, hígado y riñón. Resultados: los modelos experimentales bajo ingesta de agua de mar con relación al grupo sin agua de mar, independiente del tipo de anemia inducida, evidenciaron diferencias estadísticamente significativas (p< 0,05) con restablecimiento de los parámetros hematológicos. El bazo se observó en tamaño y condiciones normales evidenciando restablecimiento del daño inmunológico ocasionado. Órganos como riñón e hígado no mostraron daños morfológicos. Conclusiones: el agua de mar isotónica resultó útil como terapia complementaria en los modelos experimentales, en casos de anemias causadas por dietas pobres en proteínas, vitaminas y minerales, y desencadenadas por medicamentos. MÉD.UIS. 2014;27(3):9-18.

Introduction: the anaemia is a major public health worldwide, according to the World Health Organization worldwide affects 1620 million people in Latin America and the Caribbean 31% of the population has problems anemia's associated with deficient diets iron and related malnutrition. Objective: evaluating those therapeutic effects of isotonic seawater microfiltered to cold as complementary therapy in biological models with experimentally induced anaemia by poor diets in protein, vitamins and minerals, and by medicaments. Materials and Methods: they counted for eight weeks with 50 NMRI mice of both genders, adult - young age. Were divided into four experimental groups of 10 mice each, including a control group. Evaluated to the beginning and end of the experiment were the hematological parameters, salivary pH, copper and zinc. At the end of the experiment they were sacrificed by cervical dislocation and autopsied stops examine spleen, liver and kidney. Results: the experimental models under seawater intake relative to the group without sea water, independent of the type of induced anaemia, showed statistically significant differences (p < 0.05) with restoration of hematological parameters. The spleen was observed in size and showing restoration of normal immune damage caused. Organs such as kidney and liver showed no morphological damage. Conclusions: isotonic seawater useful as complementary therapy in experimental models, in cases of anemia caused by diets low in protein, vitamins and minerals, and triggered by medicament. MÉD. UIS. 2014;27(3):9-18.

Caracterização da capacidade funcional, nível cognitivo e força muscular respiratória de idosas com síndrome parkinsoniana / Characterization of functional capacity, cognitive level and respiratory muscle strenght of elderly women with parkinsonian syndrome

Este estudo teve como objetivo caracterizar uma população idosa asilar com síndrome parkinsoniana quanto a forca muscular respiratória, nível cognitivo e independência nas atividades de vida diária. O estudo caracterizou-se como uma serie de casos prospectivos, na qual a amostra foi constituída por sete idosas diagnosticadas com síndrome parkinsoniana. O estado cognitivo das participantes foi avaliado por meio do Mini-Exame de Estado Mental, a capacidade funcional pelo Índice de Katz e a forca muscular respiratória (inspiratória e expiratória) por manovacuometria. Os resultados deste estudo demonstraram que, em relação ao nível cognitivo, 71,44% das idosas apresentaram perda cognitiva moderada; na capacidade funcional, a maioria apresentou independência parcial (57,14%) e a forca muscular respiratória se apresentou abaixo do esperado. Através dos resultados, foi possível concluir que, alem do próprio envelhecimento, a SP pode ter influenciado de maneira negativa os aspectos cognitivos, a forca muscular respiratória e a capacidade funcional das idosas avaliadas.

This study aim was to characterize an asylum elderly population with Parkinsonian Syndrome regarding their respiratory muscle strength, cognitive level and independence in activities of daily living. The study was characterized as a series of prospective cases, in which the sample was composed of seven elderly women diagnosed with Parkinsonian Syndrome. The cognitive status of the participants was assessed through the Mini-Mental State Examination, the functional capacity by the Katz Index and the respiratory muscle strength (inspiratory and expiratory) by manovacuometry. The results of this study showed that in relation to the cognitive level, 71.44% of the elderly had mild cognitive impairment; in functional capacity, the majority showed partial independence (57.14%) and the respiratory muscle strength performed below expectations. Through the results, one might conclude that, in addition to aging itself, the PS may have influenced negatively the cognitive aspects, the respiratory muscle strength and the functional capacity of the assessed elderly.