Development and validation of a one-tube, nested real-time PCR method suitable for routine detection of Mycobacterium bovis in animal tissue.

AIMS: Development and validation of a real-time PCR test for high-throughput routine screening of animal tissue for Mycobacterium bovis and other Mycobacterium tuberculosis complex (MTBC) members. METHODS AND RESULTS: A preliminary study compared the results of a combination of five tissue preparation/DNA extraction methods and nine PCR assays on a panel of 92 cattle tissue samples of known M. bovis culture status (55 positive and 37 negative). The combination of DNA extraction and PCR was found to be important in achieving optimal detection of M. bovis. The optimal combination of a simple tissue preparation/DNA extraction method and a one-tube, nested real-time PCR to maximize the sensitivity of detection of an M. bovis-specific RD4 deletion and an IS1081 MTBC-specific target was selected for further evaluation. In total, tissue samples collected from 981 cattle and 366 non-bovine animals and submitted for routine TB culture were parallel tested with the selected method, as well as tissue samples obtained from 156 animals in certified TB-free cattle herds. CONCLUSION: For cattle, the optimized RD4-IS1081 PCR test exhibited a diagnostic sensitivity of 96% (95% CI: 94-97%) and specificity of 97% (95% CI: 95-98%) compared to culture. Specificity was 100% when testing the 156 samples from known TB-free cattle. For non-bovine species, the PCR had a diagnostic sensitivity of 93% (95% CI: 83-98%) and a specificity of 99% (95% CI: 97-100%).

Effects of orlistat on visceral fat after liposuction.

BACKGROUND: Liposuction can aggravate metabolic complications associated with obesity. It has been shown that the recovery of weight lost through these interventions is associated with body fat redistribution toward the visceral cavity, increasing metabolic risk factors for coronary heart disease such as insulin resistance and high triglyceride levels. OBJECTIVES: The aim of this study was to evaluate the consequences of liposuction on body mass redistribution and metabolic parameters 6 months after surgery and to evaluate the use of orlistat treatment (tetrahydrolipstatin) in controlling these parameters. METHODS: A population of 31 women with a mean body mass index of 26.17+/-3.9 kg/m(2) and undergoing liposuction of more than 1,000 cm(3), was studied. Twelve of them were treated postsurgery with 120 mg of orlistat every 8 hours for the following 6 months. Anthropometric, analytical, and radiological (computed tomography) tests were performed to quantify visceral fat area before surgery and 6 months after surgery. RESULTS: Despite weight loss after liposuction, visceral fat was not modified. Patients treated with orlistat showed a greater reduction in visceral fat, although not statistically significant. Orlistat use induced a reduction in low-density lipoprotein cholesterol values of 20.0+/-22.5 mg/dL, compared with an increase of 8.46+/-20.1 mg/dL in controls (p=.07). CONCLUSIONS: Visceral fat does not decrease despite weight loss after liposuction. Orlistat use postliposuction might be a useful tool because it shows a tendency to reduce visceral fat and improve blood lipids profile.

[Virilization of a post-menopausal woman. Diagnostic and therapeutic considerations]. / Virilización en una mujer posmenopáusica. Consideraciones diagnósticas y terapéuticas.

Virilizing tumors are rare disorders that commonly develop in the adrenal cortex or the ovaries. Because these tumors are usually millimetric, androgen-secreting ovarian tumors can be particularly difficult to identify through commonly available imaging techniques and can thus pose a diagnostic challenge. We present the case of a 71-year-old woman who presented with severe clinical hyperandrogenism and biochemical traits suggestive of an ovarian source but who was diagnosed with a 10-cm adrenal mass.

Carbon monoxide exposure in households in Ciudad Juárez, México.

This study assessed exposure to carbon monoxide from gas and wood heater emissions in a sample of 64 households in peri-urban residential areas in Ciudad Juárez, Chihuahua, México. Indoor and outdoor carbon monoxide concentrations and temperatures were monitored for a continuous period of 1 week at 1 and 6-min intervals, respectively. The moving average carbon monoxide concentrations were compared to the World Health Organization (WHO) standards for carbon monoxide. Sixty-seven percent of households with gas heaters and 60% of households with wood heaters exceeded a health-based standard at some point during the monitoring. The difference between indoor and outdoor temperatures was modestly correlated with average carbon monoxide exposure (r=0.35, p-value <0.01). Heater type may be a stronger determinant of exposure, as households with a particular heater model (the El Sol FM-210) were significantly more likely to be among the more highly exposed households (odds ratio of 4.8, p-value of 0.02). A variety of health effects were pooled and found at elevated frequency in the households that exceeded the 8-h standard of 9ppm (odds ratio=5.1, p-value=0.031). These results highlight the need for further efforts to identify and mitigate potentially hazardous carbon monoxide exposures, particularly in moderate-income countries with cooler climates.

Evaluation of VOC concentrations in indoor and outdoor microenvironments at near-road schools.

A 14-week air quality study, characterizing the indoor and outdoor concentrations of 18 VOCs at four El Paso, Texas elementary schools, was conducted in Spring 2010. Three schools were in an area of high traffic density and the fourth school, considered as a background school, was situated in an area affected minimally by stationary and mobile sources of air pollution. Passive samplers were deployed for monitoring and analyzed by GC/MS. Differences in the concentration profiles of the BTEX species between the high and low traffic density schools confirmed the pre-defined exposure patterns. Toluene was the predominant compound within the BTEX group and the 96-hr average outdoor concentrations varied from 1.16 to 4.25 µg/m3 across the four schools. Outdoor BTEX species were strongly correlated with each other (0.63 < r < 1.00, p < 0.05) suggesting a common source: vehicular traffic emissions. As expected, the strength of the associations between these compounds was more intense at each of the three high-exposure schools in contrast to the low-exposure school. This was further corroborated by the results obtained from the BTEX inter-species ratios (toluene: benzene and m, p- xylenes: ethylbenzene). Certain episodic events during the study period resulted in very elevated concentrations of some VOCs such as n-pentane. Indoor concentration of compounds with known indoor sources such as α -pinene, d-limonene, p-dichlorobenzene, and chloroform were generally higher than their corresponding outdoor concentrations. Cleaning agents, furniture polishes, materials used in arts and crafts activities, hot-water usage, and deodorizing cakes used in urinal pots were the likely major sources for these high indoor concentrations. Finally, retrospective assessment of average ambient BTEX concentrations over the last twenty years suggest a gradual decrement in this border region.

Assessment of indoor and outdoor PM species at schools and residences in a high-altitude Ecuadorian urban center.

An air monitoring campaign to assess children's environmental exposures in schools and residences, both indoors and outdoors, was conducted in 2010 in three low-income neighborhoods in Z1 (north), Z2 (central), and Z3 (southeast) zones of Quito, Ecuador - a major urban center of 2.2 million inhabitants situated 2850 m above sea level in a narrow mountainous basin. Z1 zone, located in northern Quito, historically experienced emissions from quarries and moderate traffic. Z2 zone was influenced by heavy traffic in contrast to Z3 zone which experienced low traffic densities. Weekly averages of PM samples were collected at schools (one in each zone) and residences (Z1 = 47, Z2 = 45, and Z3 = 41) every month, over a twelve-month period at the three zones. Indoor PM2.5 concentrations ranged from 10.6 ± 4.9 µg/m(3) (Z1 school) to 29.0 ± 30.5 µg/m(3) (Z1 residences) and outdoor PM2.5 concentrations varied from 10.9 ± 3.2 µg/m(3) (Z1 school) to 14.3 ± 10.1 µg/m(3) (Z2 residences), across the three zones. The lowest values for PM10-2.5 for indoor and outdoor microenvironments were recorded at Z2 school, 5.7 ± 2.8 µg/m(3) and 7.9 ± 2.2 µg/m(3), respectively. Outdoor school PM concentrations exhibited stronger associations with corresponding indoor values making them robust proxies for indoor exposures in naturally ventilated Quito public schools. Correlation analysis between the school and residential PM size fractions and the various pollutant and meteorological parameters from central ambient monitoring (CAM) sites suggested varying degrees of temporal relationship. Strong positive correlation was observed for outdoor PM2.5 at Z2 school and its corresponding CAM site (r = 0.77) suggesting common traffic related emissions. Spatial heterogeneity in PM2.5 concentrations between CAM network and sampled sites was assessed using Coefficient of Divergence (COD) analysis. COD values were lower when CAM sites were paired with outdoor measurements (<0.2) and higher when CAM and indoor values were compared (>0.2), suggesting that CAM network in Quito may not represent actual indoor exposures.

Applying the mental models framework to carbon monoxide risk in northern Mexico.

OBJECTIVES: Low-income residents of northern Mexico rely on unvented heaters during the winter, a practice that puts them at elevated risk for carbon monoxide intoxication. The goal of this study is to develop a communication protocol for carbon monoxide intoxication risks among the primarily low socioeconomic status population of Ciudad Juárez, Chihuahua, Mexico. METHODS: The mental models risk communication approach was used to identify important gaps in public understanding. This approach consists of step-by-step assessment of information needs and effectiveness of risk communication efforts by using interviews and surveys. RESULTS: The mental models process uncovered a key technical misunderstanding, the subject population's belief that carbon monoxide can be seen or smelled, which may result in a risk-prone behavior: failure to use a carbon monoxide detector. A communication protocol was designed to address this and other knowledge gaps, and it produced significant improvements in subjects' knowledge in a pretest/posttest evaluation. CONCLUSIONS: The mental models process was successful in developing a communication instrument capable of improving knowledge in the subject population. Future research needs include assessing the extent to which this instrument succeeds in changing behavior and reducing the risk of carbon monoxide intoxication. Future interventional efforts may focus on encouraging people to use carbon monoxide detectors.

Tomato BRASSINOSTEROID INSENSITIVE1 is required for systemin-induced root elongation in Solanum pimpinellifolium but is not essential for wound signaling.

The tomato Leu-rich repeat receptor kinase BRASSINOSTEROID INSENSITIVE1 (BRI1) has been implicated in both peptide (systemin) and steroid (brassinosteroid [BR]) hormone perception. In an attempt to dissect these signaling pathways, we show that transgenic expression of BRI1 can restore the dwarf phenotype of the tomato curl3 (cu3) mutation. Confirmation that BRI1 is involved in BR signaling is highlighted by the lack of BR binding to microsomal fractions made from cu3 mutants and the restoration of BR responsiveness following transformation with BRI1. In addition, wound and systemin responses in the cu3 mutants are functional, as assayed by proteinase inhibitor gene induction and rapid alkalinization of culture medium. However, we observed BRI1-dependent root elongation in response to systemin in Solanum pimpinellifolium. In addition, ethylene perception is required for normal systemin responses in roots. These data taken together suggest that cu3 is not defective in systemin-induced wound signaling and that systemin perception can occur via a non-BRI1 mechanism.

Patterns of Dwarf expression and brassinosteroid accumulation in tomato reveal the importance of brassinosteroid synthesis during fruit development.

Brassinosteroids (BRs) are essential for many physiological functions in plants, however little is known concerning where and when they are synthesized. This is especially true during flower and fruit production. To address this we have used a promoter-GUS reporter fusion and RT-PCR to determine the relative expression levels of the tomato Dwarf (D) gene that encodes a BR C-6 oxidase. In young seedlings GUS reporter activity was observed mainly in apical and root tissues undergoing expansion. In flowers GUS activity was observed in the pedicel joints and ovaries, whereas in fruits it was strongest during early seed development and was associated with the locular jelly and seeds. RT-PCR analysis showed that tissue-specific expression of Dwarf mRNA was consistent with that of the Dwarf:GUS fusion. In good correlation with the high local Dwarf activity, quantitative measurements of endogenous BRs indicated intense biosynthesis in developing tomato fruits, which were also found to contain high amounts of brassinolide. Grafting experiments showed the lack of BR transport indicating that BR action occurs at the site of synthesis.

Cloning the tomato curl3 gene highlights the putative dual role of the leucine-rich repeat receptor kinase tBRI1/SR160 in plant steroid hormone and peptide hormone signaling.

Brassinosteroids (BRs) are plant steroid hormones that are essential for normal plant development. To gain better understanding of the conservation of BR signaling, the partially BR-insensitive tomato mutant altered brassinolide sensitivity1 (abs1) was identified and found to be a weak allele at the curl3 (cu3) locus. BR content is increased in both of these mutants and is associated with increased expression of DWARF: The tomato homolog of the Arabidopsis Brassinosteroid Insensitive1 Leu-rich repeat (LRR) receptor-like kinase, named tBri1, was isolated using degenerate primers. Sequence analysis of tBRI1 in the mutants cu3 and abs1 revealed that cu3 is a nonsense mutant and that abs1 is a missense mutant. A comparison of BRI1 homolog sequences highlights conserved features of BRI1 sequences, with the LRRs in close proximity to the island domain showing more conservation than N-terminal LRRs. The most homologous sequences were found in the kinase and transmembrane regions. tBRI1 (SR160) also has been isolated as the putative receptor for systemin, a plant peptide hormone. This finding suggests a possible dual role for tBRI1 in steroid hormone and peptide hormone signaling.

Applying the mental models framework to carbon monoxide risk in northern Mexico / Aplicación del enfoque de los modelos mentales al riesgo por monóxido de carbono en el norte de México

OBJECTIVES: Low-income residents of northern Mexico rely on unvented heaters during the winter, a practice that puts them at elevated risk for carbon monoxide intoxication. The goal of this study is to develop a communication protocol for carbon monoxide intoxication risks among the primarily low socioeconomic status population of Ciudad Juárez, Chihuahua, Mexico. METHODS: The mental models risk communication approach was used to identify important gaps in public understanding. This approach consists of step-by-step assessment of information needs and effectiveness of risk communication efforts by using interviews and surveys. RESULTS: The mental models process uncovered a key technical misunderstanding, the subject population's belief that carbon monoxide can be seen or smelled, which may result in a risk-prone behavior: failure to use a carbon monoxide detector. A communication protocol was designed to address this and other knowledge gaps, and it produced significant improvements in subjects' knowledge in a pretest/posttest evaluation. CONCLUSIONS: The mental models process was successful in developing a communication instrument capable of improving knowledge in the subject population. Future research needs include assessing the extent to which this instrument succeeds in changing behavior and reducing the risk of carbon monoxide intoxication. Future interventional efforts may focus on encouraging people to use carbon monoxide detectors.

OBJETIVOS: Los residentes de bajos ingresos del norte de México dependen durante el invierno de calefactores no ventilados, una práctica que los pone en mayor riesgo de intoxicación por monóxido de carbono. Se elaboró un protocolo de comunicación sobre los riesgos de intoxicación por monóxido de carbono en la población fundamentalmente de bajo nivel socioeconómico de Ciudad Juárez, Chihuahua, México. MÉTODOS: Se utilizó el enfoque de comunicación de riesgos de los modelos mentales para identificar importantes brechas en la comprensión pública. Este enfoque consiste en la evaluación paso a paso de las necesidades de información y la eficacia de los esfuerzos de comunicación de riesgos, mediante entrevistas y encuestas. RESULTADOS: El proceso basado en los modelos mentales descubrió un malentendido técnico clave: la población estudiada cree que el monóxido de carbono se puede ver u oler. Esto puede llevar a un comportamiento proclive al riesgo: no utilizar detectores de monóxido de carbono. Se diseñó un protocolo de comunicación para atender esta y otras brechas en el conocimiento que, según las evaluaciones previa y posterior, mejoró significativamente el nivel de conocimiento de las personas. CONCLUSIONES: El proceso basado en modelos mentales permitió desarrollar exitosamente un instrumento de comunicación capaz de mejorar el nivel de conocimiento en la población estudiada. Investigaciones futuras deben evaluar en qué grado este instrumento logra modificar el comportamiento y reducir el riesgo de intoxicación por monóxido de carbono. Nuevas intervenciones podrían centrarse en estimular el uso de detectores de monóxido de carbono.

Virilización en una mujer posmenopáusica. Consideraciones diagnósticas y terapéuticas / Virilization of a post-menopausal woman. Diagnostic and therapeutic considerations

Los tumores virilizantes son poco frecuentes, proceden del ovario o de la corteza adrenal. El diagnóstico etiológico es en ocasiones muy complejo debido a que los tumores ováricos secretores de andrógenos son habitualmente de tamaño milimétrico y no se detectan mediante las pruebas de imagen convencionales. Presentamos el caso de una paciente de 71 años de edad con hiperandrogenismo grave de perfil típicamente ovárico, en la que se detectó una masa adrenal de 10 cm (AU)

Virilizing tumors are rare disorders that commonly develop in the adrenal cortex or the ovaries. Because these tumors are usually millimetric, androgen-secreting ovarian tumors can be particularly difficult to identify through commonly available imaging techniques and can thus pose a diagnostic challenge. We present the case of a 71-year-old woman who presented with severe clinical hyperandrogenism and biochemical traits suggestive of an ovarian source but who was diagnosed with a 10cm adrenal mass (AU)

Evaluación de la prueba de ELISA para anticuerpos contra el antígeno micobacterial P-90 en el antígeno de tuberculosis / Evaluation of ELISA antibodies test against P-90 mycobacterial antigen in tuberculosis diagnosis

OBJETIVO: Determinar la sensibilidad y especificidad del diagnóstico serológico mediante la detección de IgA específica para tuberculosis (TB). DISEÑO: Estudio observacional, prospectivo, de casos y controles. LUGAR: Hospital Nacional Dos de Mayo, en el período de octubre de 1998 hasta marzo de 1999. MATERIALES Y MÉTODOS: Se evaluó 81 casos de tuberculosis demostrados mediante frotis y cultivo y/o biopsia positiva para bacilo de Koch (BK) y 86 controles demostrados sanos. Se utilizó la prueba de diagnóstico serológico de TB mediante la respuesta de IgA al antígeno P-90, con la prueba de enzima inmunoabsorbente (kit Kreatech EIA-TB, Amsterdam, Holanda), preparada a partir del BCG. RESULTADOS: La sensibilidad de la prueba es 96,3 por ciento, la especificidad 77,9 por ciento. CONCLUSIONES: La detección de inmunoglobulina A mediante el antígeno clase Kp-90 Im CRAC del Mycobacterium tuberculosis no es de utilidad complementaria en el diagnóstico de tuberculosis, especialmente en el gran porcentaje de pacientes que es tratado como BK negativo.