High-throughput retrieval of target sequences from complex clone libraries using CRISPRi.

The capture of metagenomic DNA in large clone libraries provides the opportunity to study microbial diversity that is inaccessible using culture-dependent methods. In this study, we harnessed nuclease-deficient Cas9 to establish a CRISPR counter-selection interruption circuit (CCIC) that can be used to retrieve target clones from complex libraries. Combining modern sequencing methods with CCIC cloning allows for rapid physical access to the genetic diversity present in natural ecosystems.

Consorcio microbiano nativo con actividad catalítica para remoción de índigo y surfactantes en agua residual industrial textil a través de una matriz de inmovilización / Microbial consortium native catalytically active for removal of indigo and surfactants in textile industry wastewater through an immobilization matrix

El siguiente estudio tuvo como objetivo aislar seis diferentes cepas bacterianas provenientes de las descargas de agua utilizadas en la tintura de hilo con colorante índigo, que tuviesen capacidad de degradación de compuestos orgánicos del tinte índigo y tres surfactantes de tipo no iónicos. Igualmente, se evaluaron diferentes medios de soporte para inmovilizar las cepas seleccionadas. Las cepas con mejor capacidad de decoloración se combinaron para conformar cuatro consorcios (I, II, III, y IV) con el fin de potenciar el proceso de decoloración, considerando que la sinergia y el complemento de actividades metabólicas de cultivos mixtos dentro de una comunidad microbiana incrementan la eficiencia de remoción de carga orgánica. Los porcentajes de remoción que se alcanzaron fueron 64, 73, 76 y 59 %, respectivamente. Los cultivos individuales no presentaron porcentajes de remoción superiores a los reportados por los consorcios, lo que permite pensar en su utilización para la remoción de tintes índigos en aguas residuales.

The aim of this study was isolate six different bacterial strains from water discharges used in dyeing yarn with indigo, capable of degradation of organic compounds with indigo dye and three type nonionic surfactants. Similarly, various supporting media were evaluated for immobilizing the selected strains. Strains with better capacity were combined to form four consortia (I, II, III, and IV) in order to enhance the bleaching process, whereas synergy and complement metabolic activities of mixed cultures within a community increase microbial removal efficiency of organic load. Removal percentages were achieved which were 64, 73, 76 and 59%, respectively. Individual cultures showed no higher than rates reported by consortia removal, which suggests in its use for the removal of indigo dyes in wastewater.

Genetic variants associated with addictive behavior in Colombianaddicted and non-addicted to heroin or cocaine / Variantes genéticas asociadas con conducta adictiva en colombianos adictos y no adictos a heroína o cocaína

Objective:Determine the prevalence and compare somegenetic markers involved in addictive behavior in a groupof addicts to derivative of coca (cocaine/crack) or heroinand a control group of non-addicted people matched forgender, age and ethnicity.Methods:A 120 addicts and 120 non-addicts Colombianmale were surveyed and genotyped for 18 polymorphismof the OPRM1, DRD2, DRD4, SLC6A3, SLC6A4, ABCB1,DβH and CYP2B6 genes. For the identification of allelesmarkers were used mini-sequencing and fragment multi-plex PCR techniques; ethnicity of cases and controls wasanalyzed with 61 AIMs.Results:The age of onset use of heroin or coca derivati-ves (cocaine/crack) was 16,5±6 years and 99,2 percent of themconsume several illicit drugs. It showed that controlsand addicts belong to the same ethnic group. Significantdifferences between addicts and controls in relation toschooling, marital status, social security family historyof substance abuse (p<0,001), Int8-VNTR SLC6A3 gene(p=0,015) and SNP 3435C>T ABCB1 gene (p=0,001) werefound.Conclusion:The present results indicate that the VNTR-6R polymorphism of the gene SLC6A3 and the genoty-pe 3435CC in the ABCB1 gene, are both associated withaddictive behavior to heroin or cocaine.

Genetic variants associated with addictive behavior in Colombian addicted and non-addicted to heroin or cocaine.

OBJECTIVE: Determine the prevalence and compare some genetic markers involved in addictive behavior in a group of addicts to derivative of coca (cocaine/crack) or heroin and a control group of non-addicted people matched for gender, age and ethnicity. METHODS: A 120 addicts and 120 non-addicts Colombian male were surveyed and genotyped for 18 polymorphism of the OPRM1, DRD2, DRD4, SLC6A3, SLC6A4, ABCB1, DßH and CYP2B6 genes. For the identification of alleles markers were used mini-sequencing and fragment multiplex PCR techniques; ethnicity of cases and controls was analyzed with 61 AIMs. RESULTS: The age of onset use of heroin or coca derivatives (cocaine/crack) was 16.5±6 years and 99.2% of them consume several illicit drugs. It showed that controls and addicts belong to the same ethnic group. Significant differences between addicts and controls in relation to schooling, marital status, social security family history of substance abuse (p <0.001), Int8-VNTR SLC6A3 gene (p= 0.015) and SNP 3435C>T ABCB1 gene (p= 0.001) were found. CONCLUSION: The present results indicate that the VNTR- 6R polymorphism of the gene SLC6A3 and the genotype 3435CC in the ABCB1 gene, are both associated with addictive behavior to heroin or cocaine.

OBJETIVO: Determinar la prevalencia y comparar marcadores genéticos involucrados en conducta adictiva en un grupo de adictos a cocaína/crack o heroína y en un grupo control de no adictos apareados por g énero, edad y etnicidad. METODOLOGÍA: 120 varones adictos y 120 no adictos fueron encuestados y genotipificados para 18 alelos de los genes OPRM1, DßH, DRD2, DRD4, SLC6A3, SLC6A4, ABCB1 y CYP2B6. Para la identificación de los diferentes alelos se utilizaron las técnicas de minisecuenciaci ón y multiplex PCR. La etnicidad de casos y controles fue analizada con 61 marcadores ancestro-informativos. RESULTADOS: La edad de inicio en el consumo de heroína o derivados de la coca fue de 16.5±6 años y el 99.2% de ellos eran policonsumidores. Los controles y los adictos pertenecen al mismo grupo étnico. Diferencias significativas entre adictos y controles fueron encontradas en relación con escolaridad (p <0.001), estado civil (p <0.001), seguridad social (p <0.001) e historia familiar de drogadicción (p <0.001), genotipo int8-VNTR del gen SLC6A3 (p= 0.015) y al SNP 3435C>T (rs1045642) del gen ABCB1 (p= 0.001). CONCLUSIÓN: Los resultados indican que el polimorfismo VNTR-6R del gen SLC6A3 y el genotipo 3435CC en el gen ABCB1, están asociados con conducta adictiva a heroína o cocaína.

Development of a panel of genome-wide ancestry informative markers to study admixture throughout the Americas.

Most individuals throughout the Americas are admixed descendants of Native American, European, and African ancestors. Complex historical factors have resulted in varying proportions of ancestral contributions between individuals within and among ethnic groups. We developed a panel of 446 ancestry informative markers (AIMs) optimized to estimate ancestral proportions in individuals and populations throughout Latin America. We used genome-wide data from 953 individuals from diverse African, European, and Native American populations to select AIMs optimized for each of the three main continental populations that form the basis of modern Latin American populations. We selected markers on the basis of locus-specific branch length to be informative, well distributed throughout the genome, capable of being genotyped on widely available commercial platforms, and applicable throughout the Americas by minimizing within-continent heterogeneity. We then validated the panel in samples from four admixed populations by comparing ancestry estimates based on the AIMs panel to estimates based on genome-wide association study (GWAS) data. The panel provided balanced discriminatory power among the three ancestral populations and accurate estimates of individual ancestry proportions (R² > 0.9 for ancestral components with significant between-subject variance). Finally, we genotyped samples from 18 populations from Latin America using the AIMs panel and estimated variability in ancestry within and between these populations. This panel and its reference genotype information will be useful resources to explore population history of admixture in Latin America and to correct for the potential effects of population stratification in admixed samples in the region.