Generation of TALEN-mediated GRdim knock-in rats by homologous recombination.

Transcription Activator-Like Effector Nucleases (TALEN) are potential tools for precise genome engineering of laboratory animals. We report the first targeted genomic integration in the rat using TALENs (Transcription Activator-Like Effector Nucleases) by homology-derived recombination (HDR). We assembled TALENs and designed a linear donor insert targeting a pA476T mutation in the rat Glucocorticoid Receptor (Nr3c1) namely GR(dim), that prevents receptor homodimerization in the mouse. TALEN mRNA and linear double-stranded donor were microinjected into rat one-cell embryos. Overall, we observed targeted genomic modifications in 17% of the offspring, indicating high TALEN cutting efficiency in rat zygotes

Contrast Enhancement of Optical Coherence Tomography Images Using Branched Gold Nanoparticles

We propose the use of branched gold nanoparticles (B-GNPs) as a contrast agent for optical coherence tomography (OCT) imaging. Our results show that even when the central source of our OCT (1325 nm) is too far from the maximum peak of the plasmon resonance, branched nanoparticles scatter light very efficiently at this wavelength. B-GNPs were tested as a contrast agent in water and agarose-TiO2 tissue phantoms; the estimated increments in contrast were 9.19 dB and 15.07 dB for branched nanoparticles in water with concentrations of 2.2×109 NPs/mL and 6.6×109 NPs/mL, respectively, while for agarose-TiO2 tissue phantoms the estimated value was 3.17 dB. These results show the promising application of B-GNPs as a contrast agent for tissue imaging using OCT, not only for sources at 1325 nm but also at other central wavelengths located between 800 and 1000 nm

Lista patrón de los tiburones, rayas y quimeras (Chondrichthyes, Elasmobranchii, Holocephali) de México

Llista patró dels taurons, rajades i quimeres (Chondrichthyes, Elasmobranchii, Holocephali) de Mèxic Es presenta una relació de les espècies de taurons, rajades  i quimeres (condrictis) que han estat registrades a les aigües territorials de Mèxic, basada en revisions de bases de dades, literatura, examen físic dels espècimens de col·leccions científiques i registres d’exemplars recol·lectats en treballs de camp durant els darrers quatre anys i que no han estat publicats. La relació conté informació de 214 espècies de condrictis que, com a mínim, han estat registrades a les aigües marines i salobroses de Mèxic, pertanyents a 84 gèneres, 40 famílies i 14 ordres. S’hi inclouen vuit espècies de quimeres, 95 de batoïdeus i 111 de taurons. La condrictiofauna mexicana és una de les més riques del món, amb el 17,3% o més de les espècies conegudes. A més a més, s’ hi inclouen 16 espècies que és probable que siguin presents a Mèxic atesos els seus patrons de distribució.Checklist of sharks, rays and chimaeras (Chondrichthyes, Elasmobranchii, Holocephali) from Mexico We present an annotated checklist of the species of sharks, rays and chimaeras (chondrichthyan fishes) occurring in Mexican waters, based on a thorough review of the literature and electronic database searches, examination of museum collection specimens, and unpublished records obtained during fieldwork conducted in the last four years. The checklist contains information of at least 214 species of chondrichthyan fishes that occur in Mexican marine and brackish waters, assigned to 84 genera, 40 families and 14 orders. It includes eight species of chimaeras, 95 batoids and 111 sharks. Condrichthyan fauna in Mexico is one of the richest in the world, with almost 17.3% of the known species. An additional  16 species are included  as their occurrence in Mexican marine waters is probable according to distributional patterns.Se presenta un listado anotado de las especies de tiburones, rayas y quimeras (Condrictios) que han sido registrados en las aguas territoriales de México, basado en revisiones de bases de datos, literatura, examinación física de los especímenes de colecciones científicas y de registros de ejemplares recolectados en campo durante los últimos cuatro años y que no han sido publicados. El listado contiene información de al menos 214 especies de condrictios que se han registrado en las aguas marinas y salobres de México, pertenecientes a 84 géneros, 40 familias y 14 órdenes. Se incluyen ocho especies de quimeras, 95 de batoideos y 111 de tiburones. La condrictiofauna mexicana es una de las más ricas a nivel mundial, con al menos 17.3% de las especies conocidas. Adicionalmente, 16 especies, se incluyen como especies probables que ocurran en México debido a sus patrones de distribución.Palabras clave: Condrictios, Elasmobranquios, Batoideos, Quimeras, México

The Physcomitrella patens unique alpha-dioxygenase participates in both developmental processes and defense responses

[Background] Plant α-dioxygenases catalyze the incorporation of molecular oxygen into polyunsaturated fatty acids leading to the formation of oxylipins. In flowering plants, two main groups of α-DOXs have been described. While the α-DOX1 isoforms are mainly involved in defense responses against microbial infection and herbivores, the α-DOX2 isoforms are mostly related to development. To gain insight into the roles played by these enzymes during land plant evolution, we performed biochemical, genetic and molecular analyses to examine the function of the single copy moss Physcomitrella patens α-DOX (Ppα-DOX) in development and defense against pathogens.[Results] Recombinant Ppα-DOX protein catalyzed the conversion of fatty acids into 2-hydroperoxy derivatives with a substrate preference for α-linolenic, linoleic and palmitic acids. Ppα-DOX is expressed during development in tips of young protonemal filaments with maximum expression levels in mitotically active undifferentiated apical cells. In leafy gametophores, Ppα-DOX is expressed in auxin producing tissues, including rhizoid and axillary hairs. Ppα-DOX transcript levels and Ppα-DOX activity increased in moss tissues infected with Botrytis cinerea or treated with Pectobacterium carotovorum elicitors. In B. cinerea infected leaves, Ppα-DOX-GUS proteins accumulated in cells surrounding infected cells, suggesting a protective mechanism. Targeted disruption of Ppα-DOX did not cause a visible developmental alteration and did not compromise the defense response. However, overexpressing Ppα-DOX, or incubating wild-type tissues with Ppα-DOX-derived oxylipins, principally the aldehyde heptadecatrienal, resulted in smaller moss colonies with less protonemal tissues, due to a reduction of caulonemal filament growth and a reduction of chloronemal cell size compared with normal tissues. In addition, Ppα-DOX overexpression and treatments with Ppα-DOX-derived oxylipins reduced cellular damage caused by elicitors of P. carotovorum.[Conclusions] Our study shows that the unique α-DOX of the primitive land plant P. patens, although apparently not crucial, participates both in development and in the defense response against pathogens, suggesting that α-DOXs from flowering plants could have originated by duplication and successive functional diversification after the divergence from bryophytes.This work was supported by Agencia Nacional de Investigación e Innovación (ANII) [grants FCE2007_376, FCE2011_6095, fellowships BE_POS_2009_726 (A. Castro) and BE_POS_2010_2533 (L. Machado)], UdelaR Uruguay/CSIC Spain (Joint project), the Swedish Research Council, and Programa de Desarrollo de las Ciencias Básicas (PEDECIBA) Uruguay. The Ppα-DOX cDNA was obtained from the RIKEN Biological Research Center, Tsukuba, Japan

Pathogenicity island cag, vacA and IS605 genotypes in Mexican strains of Helicobacter pylori associated with peptic ulcers

<p>Abstract</p> <p>Background</p> <p><it>Helicobacter pylori </it>is associated with chronic gastritis, peptic ulcers, and gastric cancer. Two major virulence factors of <it>H. pylori </it>have been described: the pathogenicity island <it>cag </it>(<it>cag </it>PAI) and the vacuolating cytotoxin gene (<it>vacA</it>). Virtually all strains have a copy of <it>vacA</it>, but its genotype varies. The <it>cag </it>PAI is a region of 32 genes in which the insertion of IS<it>605 </it>elements in its middle region has been associated with partial or total deletions of it that have generated strains with varying virulence. Accordingly, the aim of this work was to determine the <it>cag </it>PAI integrity<it>, vacA </it>genotype and IS<it>605 </it>status in groups of isolates from Mexican patients with non-peptic ulcers (NPU), non-bleeding peptic ulcers (NBPU), and bleeding peptic ulcers (BPU).</p> <p>Methods</p> <p>The <it>cag </it>PAI integrity was performed by detection of eleven targeted genes along this locus using dot blot hybridization and PCR assays. The <it>vacA </it>allelic, <it>cag </it>PAI genotype 1 and IS<it>605 </it>status were determined by PCR analysis.</p> <p>Results</p> <p>Groups of 16-17 isolates (n = 50) from two patients with NPU, NBPU, and BPU, respectively, were studied. 90% (45/50) of the isolates harbored a complete <it>cag </it>PAI. Three BPU isolates lacked the <it>cag </it>PAI, and two of the NBPU had an incomplete <it>cag </it>PAI: the first isolate was negative for three of its genes, including deletion of the <it>cagA </it>gene, whereas the second did not have the <it>cagM </it>gene. Most of the strains (76%) had the <it>vacA </it>s1b/m1 genotype; meanwhile the IS<it>605 </it>was not present within the <it>cag </it>PAI of any strain but was detected elsewhere in the genome of 8% (4/50).</p> <p>Conclusion</p> <p>The patients had highly virulent strains since the most of them possessed a complete <it>cag </it>PAI and had a <it>vacA </it>s1b/m1 genotype. All the isolates presented the <it>cag </it>PAI without any IS<it>605 </it>insertion (genotype 1). Combined <it>vacA </it>genotypes showed that 1 NPU, 2 NBPU, and 1 BPU patients (66.6%) had a mixed infection; coexistence of <it>H. pylori </it>strains with different <it>cag </it>PAI status was observed in 1 NBPU and 2 BPU (50%) of the patients, but only two of these patients (NBPU and BPU) had different <it>vacA </it>genotypes.</p

Surveillance of Candida spp Bloodstream Infections: Epidemiological Trends and Risk Factors of Death in Two Mexican Tertiary Care Hospitals

Introduction: Larger populations at risk, broader use of antibiotics and longer hospital stays have impacted on the incidence of Candida sp. bloodstream infections (CBSI).Objective: To determine clinical and epidemiologic characteristics of patients with CBSI in two tertiary care reference medical institutions in Mexico City.Design: Prospective and observational laboratory-based surveillance study conducted from 07/2008 to 06/2010.Methods: All patients with CBSI were included. Identification and antifungal susceptibility were performed using CLSI M27-A3 standard procedures. Frequencies, Mann-Whitney U test or T test were used as needed. Risk factors were determined with multivariable analysis and binary logistic regression analysis.Results: CBSI represented 3.8% of nosocomial bloodstream infections. Cumulative incidence was 2.8 per 1000 discharges (incidence rate: 0.38 per 1000 patient-days). C. albicans was the predominant species (46%), followed by C. tropicalis (26%). C. glabrata was isolated from patients with diabetes (50%), and elderly patients. Sixty-four patients (86%) received antifungals. Amphotericin-B deoxycholate (AmBD) was the most commonly used agent (66%). Overall mortality rate reached 46%, and risk factors for death were APACHE II score >= 16 (OR = 6.94, CI95% = 2.34-20.58, p<0.0001), and liver disease (OR = 186.11, CI95% = 7.61-4550.20, p = 0.001). Full susceptibility to fluconazole, AmBD and echinocandins among C. albicans, C. tropicalis, and C. parapsilosis was observed.Conclusions: the cumulative incidence rate in these centers was higher than other reports from tertiary care hospitals from Latin America. Knowledge of local epidemiologic patterns permits the design of more specific strategies for prevention and preemptive therapy of CBSI.Pfizer Inc.Salvador Zubiran Natl Inst Med Sci & Nutr, Dept Med, Mexico City, DF, MexicoHosp Escuela Tegucigalpa, Tegucigalpa, HondurasUniversidade Federal de São Paulo, Escola Paulista Med, Div Infect Dis, São Paulo, BrazilNatl Canc Inst, Div Infect Dis, Mexico City, DF, MexicoUniv Nacl Colombia, Dept Internal Med, Bogota, ColombiaUniv Peruana Cayetano Heredia, Dept Med, Lima, PeruHosp Vargas Caracas, Caracas, VenezuelaCtr Med Caracas, Caracas, VenezuelaUniv Fed Rio de Janeiro, Univ Hosp, Rio de Janeiro, BrazilUniv Texas Med Sch Houston, Mem Hermann Texas Med Ctr, Dept Med, Houston, TX USAUniv Fed Parana, Hosp Clin, BR-80060000 Curitiba, Parana, BrazilUniv Chile, Fac Med, Hosp Luis Calvo Mackenna, Dept Pediat, Santiago 7, ChileUniv Desarrollo, Clin Alemana, Dept Med, Santiago, ChileHosp Clin Jose San Martin, Infect Dis Unit, Buenos Aires, DF, ArgentinaPontificia Univ Catolica Ecuador, Fac Med, Hosp Vozandes, Quito, EcuadorUniversidade Federal de São Paulo, Escola Paulista Med, Div Infect Dis, São Paulo, BrazilPfizer Inc.: INF-168Web of Scienc

Expression of USP18 and IL2RA is increased in individuals receiving latent tuberculosis treatment with isoniazid

Background. The treatment of latent tuberculosis infection (LTBI) in individuals at risk of reactivation is essential for tuberculosis control. However, blood biomarkers associated with LTBI treatment have not been identified. Methods. Blood samples from tuberculin skin test (TST) reactive individuals were collected before and after one and six months of isoniazid (INH) therapy. Peripheral mononuclear cells (PBMC) were isolated, and an in-house interferon-γ release assay (IGRA) was performed. Expression of chemokine ligand 4 (CCL4), chemokine ligand 10 (CXCL10), chemokine ligand 11 (CXCL11), interferon alpha (IFNA), radical S-adenosyl methionine domain-containing 2 (RSAD2), ubiquitin-specific peptidase 18 (USP18), interferon-induced protein 44 (IFI44), interferon-induced protein 44 like (IFI44L), interferon-induced protein tetratricopeptide repeats 1(IFIT1), and interleukin 2 receptor subunit alpha (IL2RA) mRNA levels were assessed by qPCR before, during, and after INH treatment. Results. We observed significantly lower relative abundances of USP18, IFI44L, IFNA, and IL2RA transcripts in PBMC from IGRA-positive individuals compared to levels in IGRA-negative individuals before INH therapy. Also, relative abundance of CXCL11 was significantly lower in IGRA-positive than in IGRA-negative individuals before and after one month of INH therapy. However, the relative abundance of CCL4, CXCL10, and CXCL11 mRNA was significantly decreased and that of IL2RA and USP18 significantly increased after INH therapy, regardless of the IGRA result. Our results show that USP18, IFI44L, IFIT1, and IL2RA relative abundances increased significantly, meanwhile the relative abundance of CCL4, CXCL11, and IFNA decreased significantly after six months of INH therapy in TST-positive individuals. Conclusions. Changes in the profiles of USP18, IL2RA, IFNA, CCL4, and CXCL11 expressions during INH treatment in TST-positive individuals, regardless of IGRA status, are potential tools for monitoring latent tuberculosis treatment

Non-perturbative scalar gauge-invariant metric fluctuations from the Ponce de Leon metric in the STM theory of gravity

We study our non-perturbative formalism to describe scalar gauge-invariant metric fluctuations by extending the Ponce de Leon metric.Comment: accepted in Eur. Phys. J.

Features of language and communication in social networks elementary school students

The written language in the social network Facebook is characterized in terms of its semiotics, semantics and expressive function. The analysis is qualitative, supported by the interpretive paradigm, which is descriptive in nature, creating groups within the framework of a curricular activity in schools in the Province of Ñuble. The written language of the posts and conversations of the study participants, whose ages ranged from 10 to 12, were analyzed according to categories of hierarchical analysis converted into codes. The analysis was facilitated by the use of the MAXQDA software, which grouped the codes and provided results linked by categories according to frequency. Results indicate that the students transmit their emotions and feelings in a written, educated and informal language with oral characteristics, structured in phrases and simple sentences with unfinished words, abbreviations and jargon expressions accompanied by an emoticon that represents the communicative situation.Se caracteriza el lenguaje escrito en la red social Facebook, en cuanto a su semiótica, semántica y función expresiva. El análisis es de carácter cualitativo, sustentado bajo el paradigma interpretativo, de naturaleza descriptiva para lo cual se crearon grupos en el marco de una actividad curricular en escuelas de la Región de Ñuble, con sujetos de estudio cuyas edades fluctuaban entre los 10 y 12 años a los cuales se les analizó el lenguaje escrito de las publicaciones y conversaciones realizadas de acuerdo a categorías de análisis jerarquizadas convertidas en códigos, el análisis fue facilitado por la utilización del software MAXQDA que agrupó los códigos entregando resultados que fueron asociados por categorías de acuerdo a frecuencia, los resultados presentan estudiantes que transmiten sus emociones y sensaciones en un lenguaje escrito de carácter culto informal con rasgos orales, estructurado en frases y oraciones simples con palabras cortadas, abreviaturas y expresiones jergales acompañadas de un emoticón que representa la situación comunicativa