A new proposal for the use of the focal animal technique in buffaloes in the Eastern Amazon.

This study aims to present a proposal for using the focal animal recording technique to evaluate the welfare of buffaloes and to verify the association between each behavior and thermal comfort indices. The study was conducted in an experimental paddock located in Santarém, Pará, Brazil. A total of 10 female Murrah animals were used. The behavior of the animals was recorded during the day, with the use of three trained observers, for 72 consecutive h. Climatic variables were collected, and the Temperature-Humidity Index (THI) and the practical Buffalo Comfort Climatic Conditions Index (BCCCIp) were determined. The multivariate technique of principal components and Spearman's correlation were employed. BCCCIp and THI were outside the thermal comfort zone at different times of the day. Grazing (P) was more frequent in the coldest hours of the day, while rumination occurred at different periods, mainly during the daytime and frequently in a lying position. There was a positive correlation between idle lying behavior and average temperature-Tmed (r = 0.583; p < 0.028), THI (r = 0.432; p < 0.034), and BCCCIp (r = 0.554; p < 0.049). There was a positive correlation between grazing and Tmed (r = 0.665; p < 0.0004) and BCCCIp (r = 0.583; p < 0.036). The standing idle behavior negatively correlated with Tmed (r = -0.718; p < 0.0001), THI (r = -0.522; p < 0.008), and BCCCIp (r = -0.8076; p < 0.0008). The lying ruminating behavior had a positive correlation with Tmed (r = 0.586; p < 0.002), THI (r = 0.477; p < 0.018), and BCCCIp (r = 0.8033; p < 0.0009). Furthermore, ruminating while standing correlated positively with Tmed (r = 0.680; p < 0.0003). The adaptation of the focal animal technique, with six observers evaluating each animal for 6 min through filming, proved to be efficient in pointing out the different behaviors of buffalo raised in Eastern Amazon fields under heat stress at different times of the day.

Diseño y validación de un ensayo de minisecuenciación múltiple para detectar polimorfismos asociados con Síndrome Metabólico / Design and validation of a multiple minisequencing assay to detect polymorphisms associated with Metabolic Syndrome

Resumen Introducción: Es importante identificar los polimorfismos de interés clínico en patologías complejas como el Síndrome Metabólico. Por esto, las metodologías para su evaluación deben estar diseñadas y validadas correctamente, esto permite optimizar recursos y tiempo en la genotipificación y detección correcta de los alelos presentes en los individuos. Objetivo: Diseñar y validar una PCR múltiple, seguida de detección por minisecuenciación, para la genotipificación de ocho polimorfismos de nucleótido simple ubicados en el gen del Receptor Beta 3-Adrenérgico (rs4994 y rs4998), gen de la Apolipoproteina A5 (rs3135506 y rs2075291), gen de la Adiponectina (rs1501299 y rs2241766) y gen del Receptor Activador de la Proliferación de los Peroxisomas tipo gamma (rs1801282 y rs1800571), asociados con el síndrome metabólico. Materiales y métodos: Se diseñaron 24 cebadores para la amplificación y detección de ocho polimorfismos de nucleótido sencillo ubicados en cuatro genes candidatos a estar asociados con el síndrome metabólico, usando el software Primer3®. Dieciséis fueron diseñados para amplificar los polimorfismos y ocho para detectarlos por minisecuenciación. Las estructuras secundarias entre los cebadores se verificaron con el software Autodimer. Los polimorfismos se amplificaron simultáneamente y los fragmentos amplificados se acoplaron a las sondas diseñadas para detectar por minisecuenciación el alelo presente, por medio de bases marcadas con fluorocromos. Finalmente, los alelos fueron detectados por electroforesis capilar en un analizador genético ABI 310 y se interpretaron con el software GeneMapper®. La validación del multiplex se realizó genotipando 20 muestras de individuos, cada uno de ellos autorizó este procedimiento por medio del consentimiento informado. Resultados: Se obtuvieron los perfiles genéticos de los 20 controles genotipados, a partir de la amplificación múltiple, seguida de minisecuenciación, diseñada y validada para detectar los ocho polimorfismos. Conclusión: Se diseñó y validó un ensayo para la detección simultánea de los polimorfismos, ubicados en cuatro genes asociados con el Síndrome metabólico. Los cuales pueden ser empleados como referencia para futuros estudios poblacionales.

Abstract Introduction: It is important to identify the polymorphisms of clinical interest in complex pathologies such as Metabolic Syndrome. Therefore, the methodologies for its evaluation must be designed and validated correctly, this permits optimization of resources and time in genotyping and correct detection of the alleles present in individuals. Objective: To design and validate a multiplex PCR, followed by detection by minisequencing, for the genotyping of eight single nucleotide polymorphisms located in the Beta 3-Adrenergic Receptor gene (rs4994 and rs4998), Apolipoprotein A5 gene (rs3135506 and rs2075291), Adiponectin gene (rs1501299 and rs2241766) and gamma-type Peroxisome Proliferation Activating Receptor gene (rs1801282 and rs1800571), associated with metabolic syndrome. Materials and methods: Twenty-four primers were designed for the amplification and detection of eight single nucleotide polymorphisms located in four candidate genes to be associated with the metabolic syndrome, using the Primer3® software. Sixteen were designed to amplify the polymorphisms and eight to detect them by minisequencing. The secondary structures between the primers were verified with Autodimer software. The polymorphisms were simultaneously amplified, and the amplified fragments were coupled to probes designed to minisequence the present allele using fluorochrome-labeled bases. Finally, the alleles were detected by capillary electrophoresis using an ABI 310 genetic analyzer and analyzed with the GeneMapper® software. The validation of the multiplex was performed by genotyping 20 individual samples, each of them authorized this procedure through informed consent. Results: The genetic profiles of the 20 genotyped controls were obtained, from multiple amplification, followed by minisequencing, designed and validated to detect the eight polymorphisms. Conclusion: An essay was designed and validated for the simultaneous detection of polymorphisms, located in four genes associated with metabolic syndrome, and can used as a reference for future population studies.

Snapshot Models of Undocumented Immigration.

Accurately estimating the size of the undocumented immigrant population is a critical component of assessing the health and security risks of undocumented immigration to the United States. To provide one such estimate, we use data from the Mexican Migration Project (MMP), a study that includes samples of undocumented Mexican immigrants to the United States after their return to Mexico. Of particular interest are the departure and return dates of a sampled migrant's most recent sojourn in the United States, and the total number of such journeys undertaken by that migrant household, for these data enable the construction of data-driven undocumented immigration models. However, such data are subject to an extreme physical bias, for to be included in such a sample, a migrant must have returned to Mexico by the time of the survey, excluding those undocumented immigrants still in the United States. In our analysis, we account for this bias by jointly modeling trip timing and duration to produce the likelihood of observing the data in such a "snapshot" sample. Our analysis characterizes undocumented migration flows including single-visit migrants, repeat visitors, and "retirement" from circular migration. Starting with 1987, we apply our models to 30 annual random snapshot surveys of returned undocumented Mexican migrants accounting for undocumented Mexican migration from 1980 to 2016. Scaling to population quantities and supplementing our analysis of southern border crossings with estimates of visa overstays, we produce lower bounds on the total number of undocumented immigrants that are much larger than conventional estimates based on U.S.-based census-linked surveys, and broadly consistent with the more recent estimates reported by Fazel-Zarandi, Feinstein, and Kaplan.

Spanish Validation of the "Life Snapshot Inventory".

INTRODUCTION: The Life Snapshot Inventory (LSI) is a self-report instrument to measure the meaningful vital, personal, and social directions. It was created in the Functional Analytic Psychotherapy as a continuous evaluation of vital changes in areas of life (family, work, love, spirituality, sexuality, health, etc.). OBJECTIVE: The aim was to validate its psychometric characteristics for the first time. METHOD: This study involved 530 participants (average age 33 years), in a Spanish sample. The questionnaire has been compared with the Rosenberg Self-Esteem Scale (RSES) to obtain convergent validity. RESULTS: The results showed a high internal consistency (α = .93) and a correlation of .61, both statistically significant. The factorial analysis showed only one factor (43.56% of variance). In addition, it was sensitive to changes due to interventions, and made it possible to differentiate those people with vital problems. CONCLUSION: This questionnaire could be a helpful measure for healthcare and clinical contexts.

INTRODUCCIÓN: El Inventario de Instantánea Vital (Life Snapshot Inventory; LSI) es un instrumento de autoinforme para medir las direcciones sociales, personales y vitales significativas para el individuo. Se ha creado desde la Psicoterapia Analítica Funcional (FAP) como una evaluación continua de los cambios en diversas áreas de la vida de un individuo (familia, trabajo, amor, espiritualidad, sexualidad, salud, etc.). Objetivo: Validar por primera vez las características psicométricas de este instrumento. METODOLOGÍA: Este estudio implicó una muestra española donde participaron 530 personas (edad media 33 años). El cuestionario se ha comparado con la Escala de Autoestima de Rosenberg (RSES) para obtener validez convergente. RESULTADOS: Los resultados mostraron una alta fiabilidad como consistencia interna (α = .93) y una correlación de .61, ambas estadísticamente significativas. El análisis factorial mostró un único factor (43.56% de la varianza). Además, el instrumento fue sensible a los cambios originados por la intervención, y permitió diferenciar aquellas personas con problemas vitales. CONCLUSIÓN: Este cuestionario podría ser una medida de gran ayuda para utilizar en contextos clínicos y sanitarios.

Molecular method for the semiquantitative identification of gastrointestinal nematodes in domestic ruminants.

Standard diagnostic methods currently in use for the identification of helminth infections in ruminants are based on the morphological analysis of immature and adult stages of parasites. This paper describes a method for the semiquantitative identification of nematodes, mainly Trichostrongyloidea, at species-level resolution. The method is based on amplification and fragment analysis followed by minisequencing of the ITS-2 region (internal transcribed spacer 2) of the ribosomal DNA of parasite eggs or larvae. This method allows for the identification of seven genera (Chabertia, Cooperia, Haemonchus, Oesophagostomum, Ostertagia, Teladorsagia, and Trichostrongylus) and 12 species (Chabertia ovina, Cooperia curticei, Cooperia punctata, Cooperia oncophora/Cooperia surnabada, Haemonchus contortus, Haemonchus placei, Haemonchus longistipes, Oesophagostomum asperum, Oesophagostomum radiatum, Ostertagia ostertagi, Trichostrongylus axei, and Trichostrongylus colubriformis) of infectious nematodes of domestic ruminants. The concordance between the morphological and molecular analyses in the detection of genera ranged from 0.84 to 0.99, suggesting the proposed detection method is specific, semiquantitative, less laborious, and highly cost-efficient.

PIMA: A population informative multiplex for the Americas.

We describe an ancestry-informative autosomal SNP multiplex designed to be a small-scale, flexible panel that can complement uniparental markers in assessing the American variability (i.e. pre-Colombian) found in contemporary indigenous American populations. This study centered on choosing SNPs with the specific characteristics of: 1) extreme allele frequency differences between indigenous Americans and the African, European and East Asian population groups that contribute to present-day population variation in the Americas; 2) high informativeness-for-assignment In values; and 3) well-spaced genomic distribution and chromosomal separation from existing small-scale forensic ancestry marker sets. The resulting capillary electrophoresis SNaPshot single base extension test was named: PIMA (Population Informative Multiplex for the Americas), comprising 26 autosomal SNPs, a single X-chromosome SNP plus the amelogenin sex marker adapted for SNaPshot. PIMA complements the established 34plex forensic ancestry panel to provide a powerful and simple tool for the analysis of American populations, including those with admixed histories, commonly encountered in America. Comparing the results obtained with the combined marker panels of PIMA and 34plex to SNP data from a much larger ancestry panel allowed us to gauge their relative efficiency. PIMA+34plex gives equivalent power to the 314-SNP 'LACE' genomic ancestry control panel, while requiring a much smaller genotyping effort. The ancestry profiles and genetic structure of 22 populations spread across the American continent were estimated using PIMA+34plex data, and those estimates were contrasted with information provided by uniparental markers (mtDNA and Y-chromosome loci) for a small set of admixed individuals from Venezuela. Our results indicate that an American genetic component is efficiently detected in contemporary American populations using a small set of ancestry informative SNPs, and these co-ancestry estimates are consistent with the known history and demography of the Americas. The small scale and high population differentiation power of PIMA, particularly when combined with 34plex, provides a practical and powerful tool for genetic studies of American populations as well as forensic DNA analyses.

A Low-Cost and Simple Genetic Screening for Cystic Fibrosis Provided by the Brazilian Public Health System.

Cystic fibrosis newborn screening was implemented in Brazil by the Public Health System in 2012. Because of cost, only 1 mutation was tested - p.Phe508del. We developed a robust low-cost genetic test for screening 11 CFTR gene mutations with potential use in developing countries.

Involvement of conformational isomerism in the complexity of the crystal network of 1-(4-nitrophenyl)-1H-1,3-benzimidazole derivatives driven by C-H...A (A = NO2, Npy and π) and orthogonal Npy...NO2 and ONO...Csp2 interactions.

A detailed structural analysis of the benzimidazole nitroarenes 1-(4-nitrophenyl)-1H-1,3-benzimidazole, C13H9N3O2, (I), 1-(4-nitrophenyl)-2-phenyl-1H-1,3-benzimidazole, C19H13N3O2, (II), and 2-(3-methylphenyl)-1-(4-nitrophenyl)-1H-1,3-benzimidazole, C20H15N3O2, (III), has been performed. They are nonplanar structures whose crystal arrangement is governed by Csp2-H...A (A = NO2, Npy and π) hydrogen bonding. The inherent complexity of the supramolecular arrangements of compounds (I) (Z' = 2) and (II) (Z' = 4) into tapes, helices and sheets is the result of the additional participation of π-πNO2 and n-π* (n = O and Npy; π* = Csp2 and NNO2) interactions that contribute to the stabilization of the equi-energetic conformations adopted by each of the independent molecules in the asymmetric unit. In contrast, compound (III) (Z' = 1) is self-paired, probably due to the effect of the steric demand of the methyl group on the crystal packing. Theoretical ab initio calculations confirmed that the presence of the arene ring at the benzimidazole 2-position increases the rotational barrier of the nitrobenzene ring and also supports the electrostatic nature of the orthogonal ONO...Csp2 and Npy...NO2 interactions.