Successful Establishment of Plasmids R1 and pMV158 in a New Host Requires the Relief of the Transcriptional Repression of Their Essential rep Genes.

Although differing in size, encoded traits, host range, and replication mechanism, both narrow-host-range theta-type conjugative enterobacterial plasmid R1 and promiscuous rolling-circle-type mobilizable streptococcal plasmid pMV158 encode a transcriptional repressor protein, namely CopB in R1 and CopG in pMV158, involved in replication control. The gene encoding CopB or CopG is cotranscribed with a downstream gene that encodes the replication initiator Rep protein of the corresponding plasmid. However, whereas CopG is an auto-repressor that inhibits transcription of the entire copG-repB operon, CopB is expressed constitutively and represses a second, downstream promoter that directs transcription of repA. As a consequence of the distinct regulatory pathways implied by CopB and CopG, these repressor proteins play a different role in control of plasmid replication during the steady state: while CopB has an auxiliary role by keeping repressed the regulated promoter whenever the plasmid copy number is above a low threshold, CopG plays a primary role by acting coordinately with RNAII. Here, we have studied the role of the regulatory circuit mediated by these transcriptional repressors during the establishment of these two plasmids in a new host cell, and found that excess Cop repressor molecules in the recipient cell result in a severe decrease in the frequency and/or the velocity of appearance of transformant colonies for the cognate plasmid but not for unrelated plasmids. Using the pMV158 replicon as a model system, together with highly sensitive real-time qPCR and inverse PCR methods, we have also analyzed the effect of CopG on the kinetics of repopulation of the plasmid in Streptococcus pneumoniae. We show that, whereas in the absence of CopG pMV158 repopulation occurs mainly during the first 45 min following plasmid transfer, the presence of the transcriptional repressor in the recipient cell severely impairs the replicon repopulation and makes the plasmid replicate at approximately the same rate as the chromosome at any time after transformation, which results in maximal plasmid loss rate in the absence of selection. Overall, these findings indicate that unrepressed activity of the Cop-regulated promoter is crucial for the successful colonization of the recipient bacterial cells by the plasmid.

[Anthropometric measures of central abdominal fat and discriminant capacity for metabolic syndrome in a Spanish population]. / Índices antropométricos estimadores de la distribución adiposa abdominal y capacidad discriminante para el síndrome metabólico en población española.

INTRODUCTION: The metabolic syndrome (MS) carries an increased risk of cardiovascular disease and diabetes mellitus. Insulin resistance is probably the mechanism underlying the changes detected in lipid and carbohydrate metabolism in these patients, who have, as a common anthropometric feature, a predominantly increased abdominal fat distribution. PATIENTS AND METHODS: A total of 3316 patients were studied, of whom 63.40% were female and 36.60 male, with a mean age of 42.36±14.63 years, and a body mass index (BMI) of 32.76±6.81kg/m(2). Weight, height and waist circumference (CC) were measured using standard techniques. The waist/height (ICA) was calculated using two indicators, expressed as waist in cm divided by height in m(2), and as waist divided by height, both in cm. The prevalence of metabolic syndrome in the sample was 33.70%. In order to assess the predictive ability of BMI, ICA and CC to detect the existence of MS, receiver operating curves (ROC) were constructed and the areas under the curve (AUC) calculated for each anthropometric parameter. RESULTS: An AUC of 0.724 (95%CI: 0.706 to 0.742), P<.001, was obtained for CC, 0.709 (95%CI: 0.691 to 0.728), P<.001 for ICA with height in m(2), and 0.729 (95%CI: 0.711 to 0.747), P<.001 for ICA with height in cm, and for the BMI it was 0.680 (95%CI 0.661-0.699), P<.001. CONCLUSIONS: Anthropometric indices that assess abdominal fat distribution have a better predictive capacity for detecting MS, compared to total adiposity indicators such as BMI.

Índices antropométricos estimadores de la distribución adiposa abdominal y capacidad discriminante para el síndrome metabólico en población española / Anthropometric measures of central abdominal fat and discriminant capacity for metabolic syndrome in a Spanish population

INTRODUCCIÓN: El síndrome metabólico conlleva un aumento del riesgo de aparición de enfermedad cardiovascular y diabetes mellitus. La existencia de un estado de resistencia a la insulina es, probablemente, el mecanismo subyacente a las alteraciones del metabolismo lipídico e hidrocarbonado detectado en estos pacientes, que presentan, como característica antropométrica frecuente, una distribución adiposa de predominio abdominal. Pacientes y métodos Se estudia una muestra de 3.316 pacientes (63,40% mujeres y 36,60 varones) de 42,36 ± 14,63 años y un índice de masa corporal (IMC) de 32,76 ± 6,81 kg/m2; en todos los pacientes se determinaron peso, talla y circunferencia de cintura ( CC ) según técnicas estandarizadas; se calculó el índice cintura/altura (ICA) mediante 2 indicadores, expresado como cintura en cm dividido por la talla en m2, y como cintura dividido por la talla, ambos en cm. La prevalencia de síndrome metabólico en la muestra fue del 33,70%. Con objeto de valorar la capacidad predictiva de IMC, ICA y CC para la detección de la existencia de SM, se construyeron las curvas operador-receptor (COR) y se calcularon las áreas bajo la curva (ABC) para cada parámetro antropométrico. RESULTADOS: Se obtuvo un ABC de 0,724 (IC 95%: 0,706-0,742), p < 0,001 para CC, de 0,709 (IC 95%: 0,691-0,728), p < 0,001 para ICA, con talla en m2, y de 0,729 (IC 95%: 0,711-0,747), p < 0,001 para ICA, con talla en cm; el ABC para IMC fue de 0,680 (IC 95%: 0,661-0,699), p < 0,001. CONCLUSIONES: Los índices antropométricos que valoran la distribución adiposa abdominal presentan mejor capacidad predictiva para la detección de SM respecto a indicadores de adiposidad total como el IMC

INTRODUCTION: The metabolic syndrome (MS) carries an increased risk of cardiovascular disease and diabetes mellitus. Insulin resistance is probably the mechanism underlying the changes detected in lipid and carbohydrate metabolism in these patients, who have, as a common anthropometric feature, a predominantly increased abdominal fat distribution. PATIENTS AND METHODS: A total of 3316 patients were studied, of whom 63.40% were female and 36.60 male, with a mean age of 42.36 ± 14.63 years, and a body mass index (BMI) of 32.76 ± 6.81 kg/m2. Weight, height and waist circumference (CC) were measured using standard techniques. The waist/height (ICA) was calculated using two indicators, expressed as waist in cm divided by height in m2, and as waist divided by height, both in cm. The prevalence of metabolic syndrome in the sample was 33.70%. In order to assess the predictive ability of BMI, ICA and CC to detect the existence of MS, receiver operating curves (ROC) were constructed and the areas under the curve (AUC) calculated for each anthropometric parameter. RESULTS: An AUC of 0.724 (95%CI: 0.706 to 0.742), P < .001, was obtained for CC, 0.709 (95% CI: 0.691 to 0.728), P < .001 for ICA with height in m2, and 0.729 (95% CI: 0.711 to 0.747), P < .001 for ICA with height in cm, and for the BMI it was 0.680 (95% CI 0.661-0.699), P < .001. CONCLUSIONS: Anthropometric indices that assess abdominal fat distribution have a better predictive capacity for detecting MS, compared to total adiposity indicators such as BMI

Asociación de polimorfismos en el gen FTO con la obesidad mórbida en la población extremeña / Association of FTO gene polymorphisms and morbid obesity in the population of Extremadura (Spain)

ObjetivoSeleccionar individuos cuya obesidad mórbida (OM) se pueda atribuir preferentemente al perfil genético individual. Tras descartar pacientes con posibles síndromes monogénicos y otras enfermedades con obesidad asociada, evaluar la asociación de la variabilidad del gen FTO (asociado con la masa grasa y la obesidad), sobre la base de los polimorfismos de un solo nucleótido (SNP) con rs1861868 y rs9939609, con la predisposición heredada a padecer OM.Pacientes y métodosTras evaluar a 270 pacientes con OM instaurada antes de los 14 años, seleccionamos a 194 por su fenotipo y la historia familiar referida; se incluyó a 289 individuos controles. Se genotiparon los cambios en el SNP rs1861868 y rs9939609 del gen FTO, y comparamos sus frecuencias genotípicas y haplotípicas en ambos grupos poblacionales.ResultadosSe confirmó la asociación del alelo A del SNP rs9939609 con la obesidad grave instaurada en la infancia en la población española. Los individuos portadores del haplotipo rs1861868 G/rs9939609 A del gen FTO mostraron un incremento del riesgo (odds ratio de 3,03; intervalo de confianza del 95%: 1,74–5,27) a padecer obesidad mórbida en nuestra población.ConclusiónAnalizar las bases genéticas de la obesidad precisa una rigurosa selección de los casos. La asociación del SNP rs9939609 con la obesidad, ampliamente descrita en distintas poblaciones, se confirma en la población española. Identificamos el primer haplotipo de riesgo al analizar el SNP rs1861868 del bloque haplotípico contiguo al que contiene el primero. Es preciso estudiar en profundidad la variabilidad interindividual del gen FTO para identificar la causa de su capacidad deletérea a la enfermedad (AU)

ObjectiveTo select individuals whose morbid obesity can be attributed mainly to their individual genetic profile. After excluding patients with potential monogenic syndromes or diseases associated with obesity, we evaluated the association of the single nucleotide polymorphisms (SNPs) rs1861868 and rs9939609 of the fat-mass and obesity-associated FTO gene with an inherited predisposition to morbid obesity.Patients and methodsWe evaluated 270 patients with morbid obesity and onset before the age of 14 years and selected 194 due to their phenotypes and family history; 289 control individuals were included. The rs1861868 and rs9939609 variants, located in the FTO gene, were genotyped. Genotype and haplotype frequencies were compared between cases and controls.ResultsThe A allele of rs9939609 was associated with severe obesity starting in childhood among the Spanish population. The rs1861868 G/rs9939609 A haplotype of the FTO gene was also significantly associated with severe obesity in our population, with an odds ratio of 3.03 (95% confidence interval, 1.74–5.27).ConclusionAnalysis of the genetic basis of obesity requires rigorous selection of cases. In this study, the association of the rs9939609 SNP with obesity widely described in distinct populations was confirmed among overweight Spanish children. Genotyping rs1861868 allowed us to identify the first risk haplotype in the FTO gene, which is located in the adjacent haplotype block containing rs9939609. In-depth study of the variability of the FTO gene is essential to define its deleterious capacity (AU)

[Association of FTO gene polymorphisms and morbid obesity in the population of Extremadura (Spain)]. / Asociación de polimorfismos en el gen FTO con la obesidad mórbida en la población extremeña.

OBJECTIVE: To select individuals whose morbid obesity can be attributed mainly to their individual genetic profile. After excluding patients with potential monogenic syndromes or diseases associated with obesity, we evaluated the association of the single nucleotide polymorphisms (SNPs) rs1861868 and rs9939609 of the fat-mass and obesity-associated FTO gene with an inherited predisposition to morbid obesity. PATIENTS AND METHODS: We evaluated 270 patients with morbid obesity and onset before the age of 14 years and selected 194 due to their phenotypes and family history; 289 control individuals were included. The rs1861868 and rs9939609 variants, located in the FTO gene, were genotyped. Genotype and haplotype frequencies were compared between cases and controls. RESULTS: The A allele of rs9939609 was associated with severe obesity starting in childhood among the Spanish population. The rs1861868 G/rs9939609 A haplotype of the FTO gene was also significantly associated with severe obesity in our population, with an odds ratio of 3.03 (95% confidence interval, 1.74-5.27). CONCLUSION: Analysis of the genetic basis of obesity requires rigorous selection of cases. In this study, the association of the rs9939609 SNP with obesity widely described in distinct populations was confirmed among overweight Spanish children. Genotyping rs1861868 allowed us to identify the first risk haplotype in the FTO gene, which is located in the adjacent haplotype block containing rs9939609. In-depth study of the variability of the FTO gene is essential to define its deleterious capacity.