Minimally invasive blood sampling method for genetic studies on Gopherus tortoises

Método de extracción de sangre mínimamente invasivo para estudios genéticos en tortugas terrestres del género Gopherus La obtención de muestras de tejido de buena calidad es la primera dificultad en cualquier estudio molecular. Esto es especialmente cierto en los estudios de manejo y conservación de la fauna silvestre. En el caso de las tortugas terrestres, la fuente más habitual de ADN son las muestras de sangre obtenidas principalmente de las venas braquial y yugular por contención química, o de individuos conscientes mediante métodos de manipulación y sujeción que pueden causar estrés en el animal. Se requiere una cantidad mínima de sangre para los ensayos del PCR. A continuación, presentamos una técnica mínimamente invasiva que ha resultado eficaz para extraer pequeñas cantidades de sangre apropiadas para realizar análisis genéticos. Además, las muestras obtenidas producen una amplificación de ADN mejor que otras fuentes celulares, como las células epiteliales cloacales. Después de dos años de aplicación en tortugas terrestres silvestres, esta técnica ha demostrado ser inofensiva. Sugerimos que el muestreo de pequeñas cantidades de sangre con esta técnica podría ser útil para otro tipo de análisis, como el seguimiento fisiológico y médico.Obtaining good quality tissue samples is the first hurdle in any molecular study. This is especially true for studies involving management and conservation of wild fauna. In the case of tortoises, the most common sources of DNA are blood samples. However, only a minimal amount of blood is required for PCR assays. Samples are obtained mainly from the brachial and jugular vein after restraining the animal chemically; or from conscious individuals by severe handling methods and clamping. Herein, we present a minimally invasive technique that has proven effective for extracting small quantities of blood, suitable for genetic analyses. Furthermore, the samples obtained yielded better DNA amplification than other cell sources, such as cloacal epithelium cells. After two years of use on wild tortoises, this technique has shown to be harmless. We suggest that sampling a small amount of blood could also be useful for other types of analyses, such as physiologic and medical monitoring.Método de extracción de sangre mínimamente invasivo para estudios genéticos en tortugas terrestres del género Gopherus La obtención de muestras de tejido de buena calidad es la primera dificultad en cualquier estudio molecular. Esto es especialmente cierto en los estudios de manejo y conservación de la fauna silvestre. En el caso de las tortugas terrestres, la fuente más habitual de ADN son las muestras de sangre obtenidas principalmente de las venas braquial y yugular por contención química, o de individuos conscientes mediante métodos de manipulación y sujeción que pueden causar estrés en el animal. Se requiere una cantidad mínima de sangre para los ensayos del PCR. A continuación, presentamos una técnica mínimamente invasiva que ha resultado eficaz para extraer pequeñas cantidades de sangre apropiadas para realizar análisis genéticos. Además, las muestras obtenidas producen una amplificación de ADN mejor que otras fuentes celulares, como las células epiteliales cloacales. Después de dos años de aplicación en tortugas terrestres silvestres, esta técnica ha demostrado ser inofensiva. Sugerimos que el muestreo de pequeñas cantidades de sangre con esta técnica podría ser útil para otro tipo de análisis, como el seguimiento fisiológico y médico

Evolution after Anti-TNF Discontinuation in Patients with Inflammatory Bowel Disease: A Multicenter Long-Term Follow-Up Study

OBJECTIVES:The aims of this study were to assess the risk of relapse after discontinuation of anti-tumor necrosis factor (anti-TNF) drugs in patients with inflammatory bowel disease (IBD), to identify the factors associated with relapse, and to evaluate the overcome after retreatment with the same anti-TNF in those who relapsed.METHODS:This was a retrospective, observational, multicenter study. IBD patients who had been treated with anti-TNFs and in whom these drugs were discontinued after clinical remission was achieved were included.RESULTS:A total of 1, 055 patients were included. The incidence rate of relapse was 19% and 17% per patient-year in Crohn''s disease and ulcerative colitis patients, respectively. In both Crohn''s disease and ulcerative colitis patients in deep remission, the incidence rate of relapse was 19% per patient-year. The treatment with adalimumab vs. infliximab (hazard ratio (HR)=1.29; 95% confidence interval (CI)=1.01-1.66), elective discontinuation of anti-TNFs (HR=1.90; 95% CI=1.07-3.37) or discontinuation because of adverse events (HR=2.33; 95% CI=1.27-2.02) vs. a top-down strategy, colonic localization (HR=1.51; 95% CI=1.13-2.02) vs. ileal, and stricturing behavior (HR=1.5; 95% CI=1.09-2.05) vs. inflammatory were associated with a higher risk of relapse in Crohn''s disease patients, whereas treatment with immunomodulators after discontinuation (HR=0.67; 95% CI=0.51-0.87) and age (HR=0.98; 95% CI=0.97-0.99) were protective factors. None of the factors were predictive in ulcerative colitis patients. Retreatment of relapse with the same anti-TNF was effective (80% responded) and safe.CONCLUSIONS:The incidence rate of inflammatory bowel disease relapse after anti-TNF discontinuation is relevant. Some predictive factors of relapse after anti-TNF withdrawal have been identified. Retreatment with the same anti-TNF drug was effective and safe

Engineering the Controlled Assembly of Filamentous Injectisomes in E. coli K-12 for Protein Translocation into Mammalian Cells.

Bacterial pathogens containing type III protein secretion systems (T3SS) assemble large needle-like protein complexes in the bacterial envelope, called injectisomes, for translocation of protein effectors into host cells. The application of these molecular syringes for the injection of proteins into mammalian cells is hindered by their structural and genomic complexity, requiring multiple polypeptides encoded along with effectors in various transcriptional units (TUs) with intricate regulation. In this work, we have rationally designed the controlled expression of the filamentous injectisomes found in enteropathogenic Escherichia coli (EPEC) in the nonpathogenic strain E. coli K-12. All structural components of EPEC injectisomes, encoded in a genomic island called the locus of enterocyte effacement (LEE), were engineered in five TUs (eLEEs) excluding effectors, promoters and transcriptional regulators. These eLEEs were placed under the control of the IPTG-inducible promoter Ptac and integrated into specific chromosomal sites of E. coli K-12 using a marker-less strategy. The resulting strain, named synthetic injector E. coli (SIEC), assembles filamentous injectisomes similar to those in EPEC. SIEC injectisomes form pores in the host plasma membrane and are able to translocate T3-substrate proteins (e.g., translocated intimin receptor, Tir) into the cytoplasm of HeLa cells reproducing the phenotypes of intimate attachment and polymerization of actin-pedestals elicited by EPEC bacteria. Hence, SIEC strain allows the controlled expression of functional filamentous injectisomes for efficient translocation of proteins with T3S-signals into mammalian cells

Explaining undocumented migration to the U.S.

The expression of NK cells activation receptors was assessed by comparative study of two groups of women workers at a chemical reagents factory, located in Zapopan, Jalisco, Mexico. Twenty of them were exposed to environmental toxics identified and quantified by gas chromatography, and 20 women unexposed to toxic substances. The expression of the surface markers CD56+ and CD3+, and of the activation receptors and co-receptors on NK cells was quantified by flow cytometry. To assess the cellular damage produced by chronic exposure to the toxics, the thiobarbituric acid reacting substances (TBARS) generated and the total plasma antioxidizing capacity (TPAC) were quantified in both groups. The exposed women had been exposed at least to 12 volatile toxic compounds, benzene, benz(a)pyrene, ethylbenzene, dimethylbenz(a)anthracene, xylene, toluene, styrene, chloroform, formaldehyde, iodine, chlorine and fluorine. Significant difference between the two groups was in the proportion of CD3 lymphocytes, 72.7 10.3% in the unexposed women versus 66.8 7.9% in the exposed group (p < 0.05). The density of expression of NKG2D and NKp30 receptors was significantly higher in the unexposed women compared to the exposed group: NKG2D were 31.3 6.3 and NKp30 were 9.5 5.2 in the unexposed women and 5.14 2.9 (p < 0.01) and 4.6 1.9 (p < 0.05), respectively in the exposed women. No statistically significant differences were found in the expression of NKp80, NKp46 and 2B4 receptors. The concentration of TBARS was lower in women from the unexposed group than the corresponding data from women of the exposed group. However, no significant difference was observed in TPAC between the two groups studied. The results of this preliminary study suggest that from the five activation receptors and co-receptors of NK cells evaluated (NKp30, NKp46, NKp80, NKG2D and 2B4), only NKp30 and NKG2D receptor expression was diminished in women exposed to toxics when compared with data from unexposed women. These results suggest that the occupational exposure to mixture of toxics is one of the important factors in the diminution of the NK cell receptor expression. " 2008 Elsevier B.V. All rights reserved.",,,,,,"10.1016/j.imlet.2008.03.010",,,"http://hdl.handle.net/20.500.12104/41410","http://www.scopus.com/inward/record.url?eid=2-s2.0-44649194964&partnerID=40&md5=9d9fa2c1197abdee0e79a6227be8b458",,,,,,"2",,"Immunology Letters",,"12

New religious movements

‘New religious movements’ (NRMs) is a term used to describe minority religions that have recently become visible in a society. They have appeared throughout the world and throughout history, and they have differed from each other with respect to their beliefs, practices, and lifestyles, as well as the effects they have on their members and on society. However, they do tend to consist of converts; to have a membership that is atypical of the rest of the population; to have a charismatic leader; to change more rapidly and radically than older, more established religions; and to evoke fear, suspicion, and, not infrequently, antagonism from nonmember