Interacción familiar y estigma social en personas que viven con VIH y SIDA en Puerto Rico

This article aims to describe the manifestation of HIV stigma in the family context and how this could impact the life of people living with HIV (PLWH). The data derive from a larger phenomenological study addressing manifestations of stigma in the lives of PLWH when interacting with the health sector. Nine focus groups were carried out in 2011 with PLWH (N=67). Eight themes emerged from the qualitative analysis. For the purpose of this article, we focus on the categories related to family dynamics: the negotiation of disclosure and non-disclosure, fear of the HIV virus and family dynamics, and life as a couple. Socio demographic information showed that 53% were between 44 to 54 years old, 80% were single, 51% were male, 42% did not complete a high school diploma, 82% were unemployed. Also, 82% described themselves as religious persons and 41% had lived with HIV for 10 years or less. Qualitative results show stigma is still present in the family context. PLWH experience fear of disclosure, discrimination, avoid initiating families or couple relationships, experience physical and verbal abuse from relatives, and even separation from other family members. After more than 30 years of the ongoing HIV epidemic, stigma is still manifested by family members with detrimental social and medical implications for PLWH. Research and educational efforts should continue addressing manifestations of stigma among family members of PLWH.Este artículo tiene el propósito de describir las manifestaciones de estigma hacia el VIH en el contexto familiar y como ello pudiera impactar la vida de las personas que viven con VIH (PLWH). La data emana de un análisis de datos de una investigación fenomenológica más amplia que aborda las manifestaciones de estigma en la vida de las personas que viven con VIH en el escenario de la salud. Nueve grupos focales fueron realizados en el 2011 con PLWH (N=67). Ocho temas emergieron del análisis cualitativo. Para propósitos de este artículo, enfatizaremos en las categorías relacionadas a las dinámicas familiares: a) la negociación de revelar o no revelar la condición, el miedo al virus del VIH y las dinámicas familiares, y vida en pareja. Datos sociodemográficos demuestran que el 53% estaban entre los 44 a 54 años de edad, 80% eran soltero, 51% eran de género masculino, 42% no completó un diploma de escuela superior, y 82% estaba desempleado. Además, 82% se consideraba una persona religiosa, y 41% había vivido con VIH por 10 años o menos. Resultados cualitativos muestran que el estigma se encuentran aun presente en el contexto familiar. Las personas que viven con VIH experimentan miedo a revelar condición, discriminación, evitan iniciar relaciones familiares o vida en pareja, experimentan abuso físico y verbal de familiares. Luego de 30 años de la condición del VIH, el estigma aún se manifiesta a través de los familiares de personas que viven con VIH, teniendo implicaciones sociales y médicas detrimentales para ellos y ellas. Esfuerzos investigativos y de educación deben continuar abordando las manifestaciones del estigma entre familiares de personas que viven con el VIH.National Institute of Mental Health (3R01 MH080694-04S1), the National Institute of Drug Abuse (UCLA HA-STTP - R25 DA035692) & the National Institute on Minority Health and Health Disparities (R25MD007607)

Purification of bacterial membrane sensor kinases and biophysical methods for determination of their ligand and inhibitor interactions

This article reviews current methods for the reliable heterologous overexpression in Escherichia coli and purification of milligram quantities of bacterial membrane sensor kinase (MSK) proteins belonging to the two-component signal transduction family of integral membrane proteins. Many of these methods were developedatLeedsalongsideProfessor SteveBaldwintowhomthisreviewisdedicated.Italsoreviewstwo biophysical methods that we have adapted successfully for studies of purified MSKs and other membrane proteins – synchrotron radiation circular dichroism (SRCD) spectroscopy and analytical ultracentrifugation (AUC), both of which are non-immobilization and matrix-free methods that require no labelling strategies. Other techniques such as isothermal titration calorimetry (ITC) also share these features but generally require high concentrations of material. In common with many other biophysical techniques, both of these biophysical methods provide information regarding membrane protein conformation, oligomerization state and ligand binding, but they possess the additional advantage of providing direct assessments of whether ligand binding interactions are accompanied by conformational changes. Therefore, both methods provide a powerful means by which to identify and characterize inhibitor binding and any associated protein conformational changes, thereby contributing valuable information for future drug intervention strategies directed towards bacterial MSKs

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The design of synthetic optogenetic tools that allow precise spatiotemporal control of biological processes previously inaccessible to optogenetic control has developed rapidly over the last years. Rational design of such tools requires detailed knowledge of allosteric light signaling in natural photoreceptors. To understand allosteric communication between sensor and effector domains, characterization of all relevant signaling states is required. Here, we describe the mechanism of light-dependent DNA binding of the light-oxygen-voltage (LOV) transcription factor Aureochrome 1a from Phaeodactylum tricornutum (PtAu1a) and present crystal structures of a dark state LOV monomer and a fully light-adapted LOV dimer. In combination with hydrogen/deuterium-exchange, solution scattering data and DNA-binding experiments, our studies reveal a light-sensitive interaction between the LOV and basic region leucine zipper DNA-binding domain that together with LOV dimerization results in modulation of the DNA affinity of PtAu1a. We discuss the implications of these results for the design of synthetic LOV-based photosensors with application in optogenetics

Dual-controlled optogenetic system for the rapid down-regulation of protein levels in mammalian cells

Abstract Optogenetic switches are emerging molecular tools for studying cellular processes as they offer higher spatiotemporal and quantitative precision than classical, chemical-based switches. Light-controllable gene expression systems designed to upregulate protein expression levels meanwhile show performances superior to their chemical-based counterparts. However, systems to reduce protein levels with similar efficiency are lagging behind. Here, we present a novel two-component, blue light-responsive optogenetic OFF switch (‘Blue-OFF’), which enables a rapid and quantitative down-regulation of a protein upon illumination. Blue-OFF combines the first light responsive repressor KRAB-EL222 with the protein degradation module B-LID (blue light-inducible degradation domain) to simultaneously control gene expression and protein stability with a single wavelength. Blue-OFF thus outperforms current optogenetic systems for controlling protein levels. The system is described by a mathematical model which aids in the choice of experimental conditions such as light intensity and illumination regime to obtain the desired outcome. This approach represents an advancement of dual-controlled optogenetic systems in which multiple photosensory modules operate synergistically. As exemplified here for the control of apoptosis in mammalian cell culture, the approach opens up novel perspectives in fundamental research and applications such as tissue engineering

Bacterial Degradation of N,N-Diethyl-m-Toluamide (DEET): Cloning and Heterologous Expression of DEET Hydrolase▿

Pseudomonas putida DTB grew aerobically with N,N-diethyl-m-toluamide (DEET) as a sole carbon source, initially breaking it down into 3-methylbenzoate and diethylamine. The former was further metabolized via 3-methylcatechol and meta ring cleavage. A gene from DTB, dthA, was heterologously expressed and shown to encode the ability to hydrolyze DEET into 3-methylbenzoate and diethylamine

Título en español.

Cured-citron was desalted by boiling gently for 5 minutes, then the water was drained off the cubes and fresh water added, in which they stood for 4 hours. Finally, the water was again changed and the samples left standing for 20 more hours. The citron cubes were successfully dehydrated in a conventional hot air tray dryer at an air temperature of 108° F. During dehydration, weight of the citron is reduced by 95 percent, and the volume by 90 per cent. The dehydrated product has a bulk density of 0.26 g./ml. Dehydrated citron has a reconstitution capacity of 98 percent for at least 12 months of storage. The data obtained in this study indicates that cured citron can be successfully dehydrated, reconstituted, and candied, resulting in a product of good quality. These experiments show that the dehydration of cured citron will reduce costs of transportation and preservation.La cidra curada se desaló de la siguiente manera: Se hirvió por 5 minutos cambiándosele luego el agua; después de 4 horas se reemplazó el agua de nuevo, dejándose la cidra en ésta por 20 horas adicionales. La cidra curada puede deshidratarse satisfactoriamente usando un secador corriente de tablillas, a una temperatura de 180° F. La cidra fermentada, al deshidratarse, pierde un 95 por ciento de su peso, un 90 por ciento de su volumen y su densidad baja hasta 0.26 g. por ml. La capacidad de rehidratación al cabo de 12 meses de almacenamiento es de un 98 por ciento. Los datos obtenidos en este estudio demuestran que la cidra curada puede deshidratarse con éxito, reconstituirse y endulzarse, obteniéndose como resultado un producto de buena calidad. Además, se ha probado que la cidra deshidratada reducirá los costos de transporte y conservación

Título en español.

The variation in the chlorophyll content of citrons was determined at different stages of maturity and at different depths in the mature green fruit. Fruit at different stages of maturity (immature green, mature green, turning ripe, and ripe) was selected from a commercial farm in Adjuntas, Puerto Rico. An examination of the endocarp revealed that it was of a greenish tint in the immature green fruit, greenish yellow in the mature green, completely yellow in the turning ripe, and whitish yellow or cream in the fully ripe fruit. Spectrophotometric determinations were made at different depths of the fruit, showing that the chlorophyll content was highest in the immature green fruit, with the mature green following closely. The chlorophyll content of fruit turning ripe was substantially lower than in the first two stages of maturity but higher than in the ripe fruit. The chlorophyll content of the citron diminishes considerably from one tissue layer to another as one cuts deeper into the fruit. Nevertheless, tissues of immature and mature green fruit contain a good amount of green pigment at greater depths. Because greenness is a characteristic index of quality in candied citron, it is advisable to use immature and mature green fruit for this purpose.Las variaciones en el contenido de clorofila se determinaron en cidras de diferentes estados de madurez y en el tejido de la fruta cortado a diferentes profundidades. La fruta se obtuvo en una finca comercial de Adjuntas, Puerto Rico, siendo las muestras representativas de cuatro estados de madurez: frutas nuevas, hechas pero aún con la cáscara verde, pintonas y maduras. Al examinar el endocarpio de las diferentes frutas se encontró que las nuevas tenían el tejido de un color verdoso, las hechas pero aún con la cáscara verde de un color verde-amarillo, las pintonas de un color amarillo y las maduras de un amarillo blancuzco, casi crema. El trabajo espectrofotométrico se llevó a cabo tomando muestras representativas a diferentes niveles de profundidad en la fruta. Los resultados demostraron que las cidras nuevas tienen el contenido más elevado de clorofila siguiéndolas de cerca las frutas hechas pero aún con la cáscara verde. El contenido en la fruta pintona es muchísimo más bajo que en la nueva y verde, pero mucho más elevado que en la madura. Los datos obtenidos indican también que el contenido de clorofila disminuye según se penetra en el mesocarpio de la fruta. Sin embargo, en las capas más profundas de la fruta nueva y de la hecha pero aún con la cáscara verde se encuentra un contenido más elevado en pigmento verde. Dado el caso de que la coloración verde en la cidra curada y endulzada es un índice de calidad, es aconsejable cosechar y fermentar la fruta nueva o la fruta ya hecha pero aún de cáscara verde

Título en español.

Studies were conducted on the extraction of coconut milk. Cold water (80° to 86° F.) and hot water (190° to 200° F.) was added to the coconut pulp in different proportions by weight. Water-addition tests were made, the ratio of water to pulp varied from 0:4 to 4:4 by weight. The results indicate that a gradual increase occurs in the percentage-yield of coconut milk as the ratio of water to pulp is increased from 0:4 to 4:4. The data shows that when the coconut milk is extracted using equal weights of water and coconut pulp (4:4 ratio), the yield is 8 to 10 percent higher than when no water is added (0:4 ratio). The results obtained from the use of cold water as compared with hot water were about the same. Chemical analyses of the coconut milk show that the fat content of both extracts is very similar with either hot or cold water. The same observation was made concerning the fat content of the press-cake.Se estudió el procedimiento de extraer la leche de coco añadiendo agua fría (de 80° a 86° F.) y agua caliente (de 190° a 200° F.), en proporciones de peso de agua a pulpa que variaron de 0:4 a 4:4. Los resultados obtenidos en estas pruebas indican que el porcentaje de leche de coco recuperado aumenta gradualmente, según varíe la cantidad de agua que se añada, desde cero hasta una cantidad igual al peso de la pulpa usada. Cuando la extracción se hace usando igual peso de agua y pulpa de coco, se obtiene un rendimiento de 8 a 10 porciento mayor que el que se obtiene sin añadir agua. Se encontró también que el uso del agua caliente en la extracción no mejora el rendimiento de leche de coco. Los análisis químicos que se hicieron demostraron que la temperatura del agua, fuese ésta fría o caliente, no afectó sustancialmente el contenido de grasa en la leche de coco y en la cachipa

Título en español.

Vitamin C stability in citron slices canned in syrup showed a retention of 77 and 73% of the original amounts of 65 and 110 mg/100 g of product, respectively, during 12.7 months storage. The average vitamin C destruction was 2.4 mg/100 g/month and 1.2 mg/100 g/month respectively for the high and low level vitamin content. There is a difference in the rate of vitamin C destruction due to the packaging form. Citron fruit bar packed in boxes had after 7 months a retention of 73% of the original vitamin C content. The same product packed in cans showed a retention of 84% after 10 months of storage.Se estudió la estabilidad de la vitamina C en dulce de cidra en sirop (rebanadas) y en pasta de cidra empacada tanto en cajas como en latas. Al cabo de 12.7 meses de almacenamiento el dulce de cidra en sirop retuvo del 73 al 77% del nivel original de vitamina C. Se encontró que durante los primeros 4 meses de almacenamiento en un dulce con 110 mg. de vitamina C por 100 g. de producto, la destrucción de esta vitamina era de aproximadamente 4 mg./100 g./mes, mientras que un dulce con 65 mg./100 g. perdió vitamina C a razón de 3 mg./100 g./mes. La destrucción de vitamina C durante todo el tiempo que duro el estudio fue de 2.4 mg./100 g./mes para el dulce de alta concentración y de 1.2 mg./100 g./mes para el de baja concentración. En pasta de cidra enriquecida con vitamina C se demostró que la forma de empaque afecta la estabilidad de la vitamina. En lotes del producto preparados simultáneamente, en los cuales la única diferencia era la forma de empaque, se encontró que la pasta enlatada contenía menos vitamina (17 mg./100 g.) que empacada en cajas. Se encontró además, que aunque existía esta diferencia inicial, al cabo de 2 meses de almacenamiento el contenido de vitamina se niveló en los dos productos en un valor de 118 mg./100 g. A los 4 meses la vitamina se niveló en los dos productos en un valor de 118 mg./100 g. A los 4 meses la vitamina C en los dos productos (enlatado y en cajas) era de 113 mg./100 g. y al cabo de 5 meses era de 110 y 109 mg./100 g., respectivamente. Al finalizar el estudio de 7 meses, la pasta de cidra en caja retuvo aproximadamente el 73% del contenido de vitamina original, mientras que en latas, al finalizar 10 meses de estudio, retuvo 84%