Comparison of Commensal and Clinical Isolates for Diversity of Plasmids in Escherichia coli and Klebsiella pneumoniae.

In this study, the plasmid content of clinical and commensal strains was analyzed and compared. The replicon profile was similar in both populations, except for L, M, A/C, and N (detected only in clinical strains) and HI1 (only in commensal strains). Although I1 and F were the most frequent replicons, only IncI1, sequence type 12 (ST12) was associated with blaCMY-2 in both populations. In contrast, the widespread resistant IncF plasmids were not linked to a single epidemic plasmid.

Faecal phageome of healthy individuals: presence of antibiotic resistance genes and variations caused by ciprofloxacin treatment.

OBJECTIVES: Antimicrobial resistance genes (ARGs) can be transferred by means of mobile genetic elements, which play a critical role in the dissemination of resistance in the bacterial community. ARG transmission within mobile genetic elements has been reported in plasmids and transposons but less frequently in bacteriophages. Here, the bacteriophage fraction of seven human faecal samples was purified and deep-sequenced to detect the presence of ARGs in the phage particles. METHODS: Seven faecal samples (five from healthy individuals and two from a patient before and after receiving ciprofloxacin treatment) were used to extract phage DNA, which was purified and then sequenced in a MiSeq (Illumina). Generated reads were checked for quality and assembled, and then the generated contigs analysed with Kraken, PHASTER, VirSorter and Prokka. Some genes were also validated by quantitative PCR. RESULTS: Analysis of the purified phage DNA by Kraken identified from 4 to 266 viruses in the samples. The viral fraction corresponded mainly to the order Caudovirales, including phages from the Siphoviridae and Myoviridae families. Bacterial genes associated with antimicrobial resistance were detected in the viral DNA, as confirmed by quantitative PCR. Higher densities of ARG-carrying phage particles were observed in the post- versus pre-ciprofloxacin treatment sample. CONCLUSIONS: The finding of ARGs in phage particles supports the description of phages as mobile elements contributing to the dissemination of bacterial antibiotic resistance and suggests ciprofloxacin treatment may play a role in the release of ARG-carrying particles, thereby increasing resistance.

Detection of mcr-1 colistin resistance gene in polyclonal Escherichia coli isolates in Barcelona, Spain, 2012 to 2015.

Colistin resistance was detected in 53 of 10,011 Escherichia coli (0.5%) by prospective phenotypic testing of consecutive clinical isolates in a single hospital in Barcelona, Spain (2012-15). The mcr-1 gene was retrospectively identified by PCR and sequencing in 15 of 50 available isolates. Each isolate had a unique PFGE pattern except for two. This clonal diversity supports the hypothesis of horizontal dissemination of the mcr-1 gene in the local study population.

Postnatal Serum Insulin-Like Growth Factor I and Retinopathy of Prematurity in Latin American Infants.

PURPOSE: Identifying at-risk infants for retinopathy of prematurity (ROP) is complex in countries with emerging economies as infants that lack conventional risk factors, such as low birth weight (BW) and young gestational age (GA), still go on to develop severe ROP. Potential biomarkers, like serum insulin-like growth factor 1 (IGF-1) and slow postnatal weight gain, have been identified as good predictors for ROP in developed countries. We sought to determine the relationship between IGF-1 levels and ROP in two Latin American countries where the burden of disease is still significant. METHODS: Prospective cohort study of infants in Guadalajara, Mexico and La Plata, Argentina. Filter-paper bloodspot IGF-1 assays were performed weekly from birth until hospital discharge or 40 weeks' postmenstrual age (PMA). RESULTS: 112 infants were studied with a median BW of 1412 g (range 620 g-2390 g) and a median GA of 33 weeks (range 25-37). There was no significant difference in IGF-1 between infants who developed ROP and those who did not. CONCLUSION: Low IGF-1 was not associated with ROP in these infants. The lack of an association between ROP and IGF-1 in Latin America supports the observation that growth-based predictive models do not perform as well in this setting where more mature babies still develop severe ROP.

External trigeminal nerve stimulation for drug resistant epilepsy: A randomized controlled trial.

BACKGROUND: External trigeminal nerve stimulation (ETNS) is an emergent, non-invasive neurostimulation therapy delivered bilaterally with adhesive skin electrodes. In previous studies, ETNS was associated to a decrease in seizure frequency in patients with focal drug-resistant epilepsy (DRE). OBJECTIVE: To determine the long-term efficacy and tolerability of ETNS in patients with focal DRE. Moreover, to explore whether its efficacy depends on the epileptogenic zone (frontal or temporal), and its impact on mood, cognitive function, quality of life, and trigeminal nerve excitability. METHODS: Forty consecutive patients with frontal or temporal DRE, unsuitable for surgery, were randomized to ETNS or usual medical treatment. Participants were evaluated at 3, 6 and 12 months for efficacy, side effects, mood scales, neuropsychological tests and trigeminal nerve excitability. RESULTS: Subjects had a median of 15 seizures per month and had tried a median of 12.5 antiepileptic drugs. At 12 months, percentage of responders was 50% in ETNS group and 0% in control group. Seizure frequency in ETNS group decreased by -43.5% from baseline. Temporal epilepsy subgroup responded better than frontal epilepsy subgroup (55.56% vs. 45.45%, respectively). Median stimulation intensity was 6.2 mA. ETNS improved quality of life, but not anxiety or depression. Long-term ETNS affected neither neuropsychological function, nor trigeminal nerve excitability. No relevant adverse events were observed. CONCLUSIONS: ETNS is an effective and well-tolerated therapy for focal DRE. Patients with temporal epilepsy showed a better response than those with frontal epilepsy. Future studies with larger populations may define its role compared to other neurostimulation techniques. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that ETNS reduces seizure frequency in patients with focal DRE.

Antibiotic resistance genes in phage particles isolated from human faeces and induced from clinical bacterial isolates.

Phage particles have emerged as elements with the potential to mobilise antibiotic resistance genes (ARGs) in different environments, including the intestinal habitat. This study aimed to determine the occurrence of ARGs in phage particles present in faecal matter and induced from strains isolated from faeces. Nine ARGs (blaTEM, blaCTX-M-1 group, blaCTX-M-9 group, blaOXA-48, qnrA, qnrS, mecA, sul1 and armA) were quantified by qPCR in the phage DNA fractions of 150 faecal samples obtained from healthy individuals who had not received antibiotic treatment or travelled abroad in the 3 months prior to sample collection. On the suspicion that the detected particles originated from bacterial flora, 82 Escherichia coli and Klebsiella pneumoniae isolates possessing at least one identified ARG (blaTEM, blaCTX-M-1 group, blaCTX-M-9 group, armA, qnrA, qnrS and sul1) were isolated and their capacity to produce phage particles carrying these ARGs following induction was evaluated. Of 150 samples, 72.7% were positive for at least one ARG, with blaTEM and blaCTX-M-9 group being the most prevalent and abundant. Of the 82 isolates, 51 (62%) showed an increase in the number of copies of the respective ARG in the phage fraction following induction, with blaTEM, blaCTX-M-1 group, blaCTX-M-9 group and sul1 being the most abundant. Phages induced from the isolates were further purified and visualised using microscopy and their DNA showed ARG levels of up to 1010 gene copies/mL. This study highlights the abundance of phage particles harbouring ARGs and indicates that bacterial strains in the intestinal habitat could be source of these particles.

Prevalence of colistin resistance in clinical isolates of Enterobacteriaceae: A four-year cross-sectional study.

OBJECTIVES: The objectives were to determine the prevalence of colistin resistance in clinical isolates of Enterobacteriaceae, and to gain knowledge on the epidemiological and clinical features of the patients. METHODS: All colistin-resistant Enterobacteriaceae consecutively isolated from clinical samples in our institution from 2012 to 2015, were included in this cross-sectional study. Intrinsic-resistant species were excluded. Minimum inhibitory concentration was performed by gradient diffusion. Detection of plasmid-encoded colistin resistance genes mcr-1 and mcr-2 was performed by amplification. Epidemiological and clinical features were reviewed. RESULTS: Of 13579 Enterobacteriaceae isolates, 91 were colistin-resistant. The overall prevalence of colistin resistance was 0.67%. The rates were higher in Enterobacter cloacae (4.2%) than Escherichia coli (0.5%) and Klebsiella pneumoniae (0.4%). One third of the isolates were multi-drug resistant (MDR). While mcr-2 was not detected, mcr-1 was detected only in E. coli. Regarding these infections, 23% were community-acquired. 89% of the patients had not received colistin previously. There were no significant differences between infections caused by mcr-1 and non-mcr-1-carrying isolates. CONCLUSIONS: Colistin resistance was not restricted to MDR isolates and to clinical settings. Most patients had no record of previous administration of colistin.

PARP-1/PARP-2 double deficiency in mouse T cells results in faulty immune responses and T lymphomas.

The maintenance of T-cell homeostasis must be tightly regulated. Here, we have identified a coordinated role of Poly(ADP-ribose) polymerase-1 (PARP-1) and PARP-2 in maintaining T-lymphocyte number and function. Mice bearing a T-cell specific deficiency of PARP-2 in a PARP-1-deficient background showed defective thymocyte maturation and diminished numbers of peripheral CD4+ and CD8+ T-cells. Meanwhile, peripheral T-cell number was not affected in single PARP-1 or PARP-2-deficient mice. T-cell lymphopenia was associated with dampened in vivo immune responses to synthetic T-dependent antigens and virus, increased DNA damage and T-cell death. Moreover, double-deficiency in PARP-1/PARP-2 in T-cells led to highly aggressive T-cell lymphomas with long latency. Our findings establish a coordinated role of PARP-1 and PARP-2 in T-cell homeostasis that might impact on the development of PARP-centred therapies.

Enhancing tumor-targeting monoclonal antibodies therapy by PARP inhibitors.

Monoclonal antibodies (mAbs) have become a successful therapeutic approach in cancer. However, some patients do not achieve long-term clinical benefit and most mAbs only exert modest effects as monotherapies. Therefore, combinations with chemotherapy are currently being investigated. Emerging studies have shown a synergistic therapeutic effect of PARP inhibitors and mAbs in cancer. PARP enzymes catalytically cleave ß-NAD+ and transfer the ADP-ribose moiety to acceptor proteins, modifying their function. In here, we update recent data about the therapeutic effect of the combination of PARP inhibitors with mAbs in cancer treatment and discuss the molecular mechanisms involved in this synergy.

Neuroprotective Effects of Low-Dose Statins in the Retinal Ultrastructure of Hypercholesterolemic Rabbits.

To evaluate the pleiotropic effects to statins, we analyze the qualitative and quantitative retinal changes in hypercholesterolemic rabbits after a low-dosage statin treatment. For this purpose, New Zealand rabbits were split into three groups: control (G0; n = 10), fed a standard diet; hypercholesterolemic (G1; n = 8), fed a 0.5% cholesterol-enriched diet for 8 months; and statins (G2; n = 8), fed a 0.5% cholesterol-enriched diet for 8 months, together with the administration of statin (pravastatin or fluvastatin sodium) at a dose of 2 mg / kg / day each diet. The retinas were analyzed by transmission electron microscopy and immunohistochemistry (glial fibrillary acidic protein). The retinal thickness of nuclear and plexiform layers were quantified in semi-thin sections. The results revealed that the low-statin-treated rabbits in comparison with the hypercholesterolemic group showed: i) a more preserved structure in all retinal layers; ii) a significant reduction in retinal thickness; iii) a decrease in cell death in the nuclear-and ganglion-cell layers; iv) a reduction of hydropic degeneration in the plexiform and nerve-fiber layers; v) a preservation of astrocytes and of the retinal area occupied by them; and vi) a better-preserved retinal vascular structure. Our findings indicate that low doses of statins can prevent retinal degeneration, acting on retinal macroglia, neurons and retinal vessels, despite that hypercholesterolemia remained unchanged. Thus, the pleiotropic effects of the statins may help safeguard the retinal ultrastructure.

El código sanitario panamericano: hacia una política de salud continental / Pan american sanitary code: towards a policy of continental health

En octubre de 1999 se cumplieron 75 años de la adopción del Código Sanitario Panamericano, primer tratado de sanidad internacional en ser firmado por todas las naciones del continente americano. Este Código, que fue ratificado en La Habana, Cuba, en 1924, en ocasión de la Séptima Conferencia Sanitaria Panamericana, subrayó la contribución de la salud humana al progreso económico de los países y al intercambio comercial y simultáneamente, señaló el papel fundamental de la higiene portuaria en la promoción de la salud. En él se hacia un llamado a la cooperación entre países con la doble finalidad de fortalecer las relaciones comerciales y proteger la salud pública, tareas cuya orquestación fue delegada a la Organización Panamericana de la Salud (OPS). En honor de tan insigne acontecimiento, el Ministerio de Salud Pública de Cuba, que también cumplía para esas fechas su 90o. aniversario, celebró en La Habana del 18 al 22 de octubre una serie de actividades conmemorativas donde se insistió en la necesidad de un esfuerzo mancomunado por encontrar solución a los problemas de orden sanitario y socioeconómico que aquejan hoy en día a los países de la Región. Destacaron entre ellas una Reunión de Ministros de Salud de Iberoamérica, actividad que dió inicio a la semana conmemorativa; la II Conferencia para la Movilización de Recursos Multisectoriales, en la que participaron las autoridades cubanas de salud pública y de otros ministerios, la OPS y otras agencias de las Naciones Unidas; y el III Seminario sobre Atención Primaria, que se centró en el intercambio de experiencias en este campo en relación con la reforma del sector sanitario y de la meta de Salud para todos en el siglo XXI. Durante la semana de actividades se repasaron los sucesos históricos que llevaron a la firma del Código Sanitario y la influencia que éste ejerció en el control y la prevención de las enfermedades que prevalecían en aquel entonces en Cuba y en los demás países del continente. La Revista Panamericana de Salud Pública/Pan American Journal of Public Health publica el siguiente relato en reconocimiento de uno de los sucesos de mayor trascendencia en la historia de la salud pública de los países americanos

Influencia vestibular en la respuesta a la maniobra de Bielschowsky en parálisis del cuarto nervio craneal / Vestibular influence in the response to Bielschowsky maneuver in fourth cranial nerve palsy

Objetivo: Describir en una serie de pacientes con parálisis unilateral del cuarto nervio craneal, la diferencia de la desviación vertical en posición erecta y supina. Material y método: Se incluyeron pacientes con diagnóstico de parálisis del cuarto nervio craneal unilateral, realizando exploración oftalmológica completa así como medición de la desviación con prismas y oclusión alterna en posición erecta y en posición supina realizando la maniobra de Bielschowsky. Resultados: Se exploraron 13 pacientes con el diagnóstico antes mencionado; la mayor parte de ellos presentaron menor desviación vertical en posición supina; así mismo al realizar la maniobra de Bielschowsky, la respuesta en posición supina fue menor que la presentada en posición erecta. Conclusiones: Los reflejos vestíbulo oculares, responsable de la mayor desviación vertical al inclinar la cabeza hacia el lado parético en casos de parálisis del nervio troclear, se atenúan en posición supina. La explicación fisiológica se basa en el control de los movimientos oculares por parte de los otolitos, los cuales reciben estimulación de a cuerdo a su posición en relación a la gravedad; esta observación clínica apoya la explicación propuesta por Bielschowsky sobre la participación del aparato vestibular en uno de los principales signos clínicos de esta entidad.

Purpose. To describe vertical deviation differences in a group of patients with unilateral fourth cranial nerve palsy in erect versus supine position. Methods. We included patients with unilateral fourth cranial nerve palsy who underwent full ophthalmologic exploration and measurement of the vertical deviation with prisms and alternate occlusion in erect and supine position performing Bielschowsky´s test. Results. Thirteen patients were explored; most of them showed less vertical deviation in the supine position, the response to the Bielschowsky´s test was smaller in the supine position than in the erect patient. Conclusion. The vestibulo-ocular reflexes responsible for the larger vertical deviation when tilting the head to the paretic side, are attenuated in the supine position; the physiologic explanation of this phenomenon is based on the otholits role in the control of ocular movements; these receptors are stimulated according to their position in relation to gravity. This clinical observation supports the explanation proposed by Bielschowsky about the role of the vestibular system in one of the main clinical sings of this entity.

Grado de desviación en posición primaria de la mirada (PPM) en lesión del III y VI nervio / Degree of deviation in primary position in III and VI nerve injury

Objetivo: Evaluar las características en el grado de desviación en las lesiones del II y VI nervio, para establecer correlación entre cada una de las situaciones clínicas. Materiales y métodos: Se estudiaron expedientes de pacientes con lesión del III y VI nervio en el Hospital Nuestra Señora de La Luz, desde enero de 2000 hasta enero de 2005. En todos los pacientes se realizó examen oftalmológico completo, donde se evidenciara limitación de -4 ó mayor en III nervio, y limitación de la abducción de -4 ó mayor en VI nervio. Resultados: Se analizaron 71 expedientes, 31 con lesión del III nervio (11 con parálisis, 20 con paresia); 40 con lesión del VI nervio (16 con parálisis y 24 con paresia). En cuanto a la desviación media y la moda según el grado de limitación, se evidenció mayor desviación en la lesión del III nervio. Conclusiones: Las desviaciones horizontales encontradas cuando existe daño al III nervio son en general de mayor magnitud que las encontradas cuando existe daño al VI nervio.

Purpose: To evaluate characteristics in the amount of deviation in III and VI nerve palsies, to establish a correlation between each one of clinical situations. Methods: Retrospective study where the files of patients with the diagnosis of III or VI nerve injury in the Hospital Nuestra Señora de La Luz were reviewed from january of 2000 to january of 2005. All patients had a complete ophthalmologic evaluation, in whitch the deviation was noticed, and in those with an affection of the III nerve the limitation of the adduction went from -4 or more, and in those with de VI nerve affected, the limitation of the abduction went from -4 or more. Results: 71 patients were included, 31 with III nerve injury and 40 with VI nerve injury. Conclusions: Horizontal deviations found in III nerve palsies are larger than those found in VI nerve palsies.