Telomeric repeat-containing RNA (TERRA) and telomerase are components of telomeres during mammalian gametogenesis.

Telomeres are ribonucleoprotein structures at the end of chromosomes composed of telomeric DNA, specific-binding proteins, and noncoding RNA (TERRA). Despite their importance in preventing chromosome instability, little is known about the cross talk between these three elements during the formation of the germ line. Here, we provide evidence that both TERRA and the telomerase enzymatic subunit (TERT) are components of telomeres in mammalian germ cells. We found that TERRA colocalizes with telomeres during mammalian meiosis and that its expression progressively increases during spermatogenesis until the beginning of spermiogenesis. While both TERRA levels and distribution would be regulated in a gender-specific manner, telomere-TERT colocalization appears to be regulated based on species-specific characteristics of the telomeric structure. Moreover, we found that TERT localization at telomeres is maintained throughout spermatogenesis as a structural component without affecting telomere elongation. Our results represent the first evidence of colocalization between telomerase and telomeres during mammalian gametogenesis.

Safety, tolerability, and immunogenicity of the novel antituberculous vaccine RUTI: randomized, placebo-controlled phase II clinical trial in patients with latent tuberculosis infection.

OBJECTIVES: To evaluate the safety, tolerability and immunogenicity of three different doses (5, 25 and 50 µg) of the novel antituberculous vaccine RUTI compared to placebo in subjects with latent tuberculosis infection. METHODS AND FINDINGS: Double-blind, randomized, placebo-controlled Phase II Clinical Trial (95 patients randomized). Three different RUTI doses and placebo were tested, randomized both in HIV-positive (n = 47) and HIV-negative subjects (n = 48), after completion of one month isoniazid (INH) pre-vaccination. Each subject received two vaccine administrations, 28 Days apart. Five patients withdrew and 90 patients completed the study. Assessment of safety showed no deaths during study. Two subjects had serious adverse events one had a retinal detachment while taking INH and was not randomized and the other had a severe local injection site abscess on each arm and was hospitalized; causality was assessed as very likely and by the end of the study the outcome had resolved. All the patients except 5 (21%) patients of the placebo group (3 HIV+ and 2 HIV-) reported at least one adverse event (AE) during the study. The most frequently occurring AEs among RUTI recipients were (% in HIV+/-): injection site reactions [erythema (91/92), induration (94/92), local nodules (46/25), local pain (66/75), sterile abscess (6/6), swelling (74/83), ulcer (20/11), headache (17/22) and nasopharyngitis (20/5)]. These events were mostly mild and well tolerated. Overall, a polyantigenic response was observed, which differed by HIV- status. The best polyantigenic response was obtained when administrating 25 µg RUTI, especially in HIV-positive subjects which was not increased after the second inoculation. CONCLUSION: This Phase II clinical trial demonstrates reasonable tolerability of RUTI. The immunogenicity profile of RUTI vaccine in LTBI subjects, even being variable among groups, allows us considering one single injection of one of the highest doses in future trials, preceded by an extended safety clinical phase. TRIAL REGISTRATION: ClinicalTrials.gov NCT01136161.

¿Cómo debe diagnosticarse el trastorno de conducta del sueño REM? / No disponible

El trastorno de conducta del sueño REM (TCSR) es una parasomnia caracterizada por excesiva actividad electromiográfica (EMG) durante el sueño REM asociada a movimientos o conductas involuntarias que pueden ser vigorosas y causar lesiones en el paciente o en el compañero de cama. El TCSR puede ser idiopático o secundario a una enfermedad neurodegenerativa que curse con depósitos de la proteína alfa-sinucleína, como la enfermedad de Parkinson. El TCSR idiopático es, en realidad, y en la mayoría de los casos, síntoma de una sinucleinopatía. Su diagnóstico correcto y preciso es importante. Actualmente, la realización de un polisomnograma (PSG) para el diagnóstico del TCSR es obligatoria, pero los criterios de la Clasificación Internacional de las Alteraciones del Sueño (ICSD-2) que indican cómo hacer el diagnóstico son inespecíficos. Por una parte, la definición actual incluye la necesidad de encontrar una "excesiva" actividad EMG fásica o tónica, pero por otra parte, no explica qué significa excesiva ni tampoco cuáles son los valores de normalidad ni los músculos que deberían estudiarse en pacientes con sospecha clínica de TCSR. En busca de respuesta a estas preguntas, aparecieron las primeras investigaciones realizadas por científicos del grupo SINBAR (Sleep Insbruck Barcelona). En su primer trabajo estudiaron cuál era el montaje EMG más apropiado para la detección de la actividad fásica que ocurre en el TCSR. Escogieron un grupo de pacientes con TCSR y les realizaron un PSG que incluía 13 músculos diferentes en todo el cuerpo y codificaron la actividad fásica presente en cada uno de ellos en miniépocas de 3 segundos. Su conclusión fue que el mejor montaje para el estudio del TCSR es el que incluye los músculos del mentón, flexor superficial de los dedos en los antebrazos y extensor corto de los dedos en las piernas. En un segundo trabajo quedó demostrado que este montaje registraba el 95 etc) y vocales (hablar, gritar, etc) ocurridas durante el REM en pacientes con TCSR. Por último, en un tercer trabajo, el grupo SINBAR concluyó que el registro de la actividad EMG del mentón ,combinada con la de los flexores superficiales de los dedos, era suficientemente específica para diferenciar entre pacientes y controles, con un punto de corte del 32TCSR de manera fácil y segura registrando solo la actividad EMG del mentón y la de los flexores superficiales de los dedos, siempre que observemos actividad en más del 32 (AU)

No disponible

Virulence and antimicrobial resistance profiles among Escherichia coli strains isolated from human and animal wastewater.

To gain insight into whether Escherichia coli isolated from humans and resistant to some common antimicrobial agents are derived from animals, 85 E. coli strains were selected by ERIC-PCR from human and animal wastewater samples. Phylogroup, pathogenicity islands (PAIs), resistance to quinolones, fluoroquinolones and presence of extended-spectrum beta-lactamases (ESBLs) were analyzed. Among the total, 55% were resistant to nalidixic acid and 38% to ciprofloxacin; 12% produced ESBLs. Chicken-derived strains were associated with quinolone and fluoroquinolone resistance and presence of ESBLs, while human strains were associated with susceptibility. Group B2 E. coli strains were associated with human origin, susceptibility to fluoroquinolones and presence of PAIs, whereas groups A, B1 and D showed a low virulence profile and a high level of antimicrobial resistance. In both human and animal wastewater, E. coli A, B1 and D were prevalent, and strains from both origins showed a similar virulence profile in each phylogroup. These findings led us to hypothesize that abusive antibiotic use in food animal production may promote the development of resistance among these intestinal E. coli phylogroups, which could later be transmitted to humans through the food supply. The low prevalence of E. coli group B2 in the animal gut may explain, at least in part, the absence of emergence of resistant B2 isolates.

Quinolone, fluoroquinolone and trimethoprim/sulfamethoxazole resistance in relation to virulence determinants and phylogenetic background among uropathogenic Escherichia coli.

INTRODUCTION: The goal of this study was to assess how resistance to quinolones, fluoroquinolones and trimethoprim/sulfamethoxazole relates to the virulence potential and phylogenetic background of clinical Escherichia coli isolates. METHODS: Among 150 uropathogens (21% resistant to quinolones, 12% resistant to fluoroquinolones and 29.3% resistant to trimethoprim/sulfamethoxazole), E. coli phylogenetic group, 15 virulence-associated genes and 7 O antigens were analysed. Clonal group A (CGA) and genomic PCR profiles were studied among trimethoprim/sulfamethoxazole-resistant isolates. RESULTS: Isolates susceptible to the three antimicrobial agents were significantly associated with phylogenetic group B2, whereas resistant isolates exhibited shifts to non-B2 groups (quinolone and fluoroquinolone-resistant isolates to group A; trimethoprim/sulfamethoxazole-resistant isolates to group D). Diverse virulence traits, including UTI-associated O antigens, were significantly less frequent among resistant isolates, particularly those resistant to fluoroquinolones (median score, 3.9 virulence factors/strain) and also to quinolones (5.2) or trimethoprim/sulfamethoxazole (6.4), as compared with the corresponding drug-susceptible isolates (median scores of 7.9, 8.6 and 7.9, respectively). Among 44 trimethoprim/sulfamethoxazole-resistant isolates, 3 (6.8%) belonged to CGA. All these 3 CGA strains caused pyelonephritis (P=0.02) and exhibited the consensus virulence profile of previously described CGA strains from abroad. CONCLUSIONS: E. coli isolates resistant to quinolones, trimethoprim/sulfamethoxazole and especially fluoroquinolones were associated with reductions in virulence traits and shifts to non-B2 phylogenetic groups. Moreover, fluoroquinolone resistance usually occurred in low-virulence E. coli group A isolates rather than in isolates from groups B2 and D which had lost virulence traits. CGA accounted for 23% of trimethoprim/sulfamethoxazole-resistant E. coli producing pyelonephritis.

First detection of a carbapenem-hydrolyzing metalloenzyme in two enterobacteriaceae isolates in Spain.

Two strains of Enterobacteriaceae, Escherichia coli and Klebsiella pneumoniae, producing VIM-1 were isolated for the first time in Spain. In both strains, bla(VIM-1) was found to be carried on a gene cassette inserted into a class 1 integron. The bla(VIM-1)-containing integron was located on a transferable plasmid.

Prevalence of clinical isolates of Escherichia coli producing inhibitor-resistant beta-lactamases at a University Hospital in Barcelona, Spain, over a 3-year period.

About 7% of 7,252 nonduplicated clinical Escherichia coli strains from a Spanish hospital showed reduced susceptibility to amoxicillin-clavulanate. Of these, 0.37% produced the IRTs TEM-30, TEM-31, TEM-33, TEM-34, TEM-37, TEM-40, TEM-51, and TEM-54; 5.3% were probable class C beta-lactamase overproducers; 0.8% were probable TEM-1 hyperproducers; 0.18% produced OXA-30; 0.15% overexpressed SHV-1; and 0.03% produced a PSE-1 enzyme.

[Structure and function of integrons]. / Estructura y función de los integrones.

Integrons are genetic elements known for their role in the acquisition and expression of genes conferring antibiotic resistance. Integrons have an integrase gene (intI), an attachment site (attI), into which individual resistance genes are inserted and a promotor sequence (Pant), allowing expression of resistance genes (cassette-associated genes), which do not have promotors. Integrase recognizes 59-be, a specific sequence in certain resistance genes, which is captured by recombination at the attI attachment site. The fragment intI - attI is highly conserved in all integrons and is called 59 -CS. Integrons have been classified according to the sequence of their integrase and the ones most frequently detected in isolated clinical strains belong to Class I. Class I integrons contain the 59 -CS region followed by gene cassettes in a variable region and finally, a conserved region known as 39 -CS containing two genes, the quaternary ammonium resistance gene (qacEDI) and the sulphonamide resistance gene (sul1); both genes are fixed in this structure. Accordingly, the structure of a Class 1 integron would be IntI - attI [R11 R21.] - qacED1 - sul1. Integrons are probably not mobile, but they are often found in transposons within conjunctive plasmids, which assures their mobility, as can be seen by their wide diffusion among bacteria.

Estructura y función de los integrones / Structure and function of integrons

Los integrones son unas piezas genéticas que han despertado gran interés porque algunos de ellos vehiculan genes de resistencia a los antimicrobianos. Desde el extremo 5´ al 3´ están formados por un fragmento que codifica una integrasa (intI) y a continuación una secuencia attI a la que se unen los genes de resistencia. Dentro de intI, en su extremo 3´ , hay una secuencia promotora Pant a partir de la cual se transcriben los genes de resistencia integrados, ya que estos genes carecen de promotor. En efecto, la integrasa reconoce en ciertos genes de resistencia (denominados genes casete) una secuencia específica denominada 59-be, que une, por recombinación, a la secuencia attI del integrón. El fragmento formado por intI-attI está altamente conservado en todos los integrones y se denomina 5 -CS. Los integrones se han clasificado según la secuencia de su integrasa. Los detectados con más frecuencia en cepas aisladas en clínica pertenecen a la clase 1. Los integrones de la clase 1 están formados por el 5 -CS y a continuación se sitúan los diferentes genes casetes captados, por lo que constituye una zona variable y, finalmente, hay una zona conservada denominada 3 -CS formada por 2 genes uno de resistencia a compuestos de amonio cuaternario (qacE I) y otro a sulfamidas (sul1); estos 2 genes, no son casetes y, por lo tanto, no son móviles sino fijos. La estructura de uno de estos integrones vendría representada por IntI-attI [R1 R2 ...]-qacE 1-sul1.Probablemente, los integrones no son móviles por sí mismos, pero con frecuencia se hallan en transposones que a su vez se encuentran en plásmidos conjugativos, por lo que su movilidad horizontal está asegurada, como se constata por su amplia difusión entre las bacterias (AU)

Novel complex sul1-type integron in Escherichia coli carrying bla(CTX-M-9).

For the present report, a novel complex class 1 integron, In60, was characterized. Part of this integron includes the bla(CTX-M-9) gene and its downstream nucleotide sequence, which shares 81% and 78% nucleotide identity with those of kluA-1 beta-lactamase and orf3 of K. ascorbata, respectively. Furthermore, a new insertion sequence, IS3000, has been found in In60. PCR analysis indicates that integron In60 is present in 33 of 34 nonclonal enterobacterial isolates carrying the putative beta-lactamase CTX-M-9.

Beta-lactamases involved in resistance to broad-spectrum cephalosporins in Escherichia coli and Klebsiella spp. clinical isolates collected between 1994 and 1996, in Barcelona (Spain).

The aim of this study was to evaluate the incidence of decreased susceptibility to broad-spectrum cephalosporins in Enterobacteriaceae that lack inducible chromosomal bla genes, and to determine the enzymes responsible for resistance. From all clinically relevant Enterobacteriaceae strains isolated between 1994 and 1996, 88 of 7054 Escherichia coli, seven of 581 Klebsiella pneumoniae and 23 of 166 Klebsiella oxytoca strains were studied because of their decreased susceptibilities to broad-spectrum cephalosporins (as reflected in intermediate susceptibilities and/or positive synergy tests and/or irregular crenellated inhibition zones). The most frequent mechanism implicated in decreased susceptibility to broad-spectrum cephalosporins displayed by E. coli and K. oxytoca was hyperproduction of chromosomal beta-lactamase, followed by plasmid-mediated SHV-1 hyperproduction in E. coli. In our hospital, the incidence of plasmid-mediated extended-spectrum beta-lactamases (ESBLs) between 1994 and 1996 was low. ESBLs were found in only 10 (0.14%) E. coli strains (six CTX-M-9, two TEM-12 and two SHV-2), in one (0.17%) K. pneumoniae strain (SHV-2) and in no K. oxytoca strains. The relatively wide variety of beta-lactamases that were detected among these common bacteria isolated from a single medical centre, including non-TEM- and non-SHV-derived ESBLs, appears epidemiologically remarkable.