UFR2709, an Antagonist of Nicotinic Acetylcholine Receptors, Delays the Acquisition and Reduces Long-Term Ethanol Intake in Alcohol-Preferring UChB Bibulous Rats.

Alcoholism is a worldwide public health problem with high economic cost and which affects health and social behavior. It is estimated that alcoholism kills 3 million people globally, while in Chile it is responsible for around 9 thousand deaths per year. Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels expressed in the central nervous system, and they were suggested to modulate the ethanol mechanism involved in abuse and dependence. Previous work demonstrated a short-term treatment with UFR2709, a nAChRs antagonist, which reduced ethanol intake using a two-bottle free-choice paradigm in University of Chile bibulous (UChB) rats. Here, we present evidence of the UFR2709 efficacy in reducing the acquisition and long-term ethanol consumption. Our results show that UFR2709 (2.5 mg/kg i.p.) reduces the seek behavior and ethanol intake, even when the drug administration was stopped, and induced a reduction in the overall ethanol intake by around 55%. Using naïve UChB bibulous rats, we demonstrate that UFR2709 could delay and reduce the genetically adaptive impulse to seek and drink ethanol and prevent its excessive intake.

Chimeric RNA: DNA TracrRNA Improves Homology-Directed Repair In Vitro and In Vivo.

Nearly 90% of human pathogenic mutations are caused by small genetic variations, and methods to correct these errors efficiently are critically important. One way to make small DNA changes is providing a single-stranded oligo deoxynucleotide (ssODN) containing an alteration coupled with a targeted double-strand break (DSB) at the target locus in the genome. Coupling an ssODN donor with a CRISPR-Cas9-mediated DSB is one of the most streamlined approaches to introduce small changes. However, in many systems, this approach is inefficient and introduces imprecise repair at the genetic junctions. We herein report a technology that uses spatiotemporal localization of an ssODN with CRISPR-Cas9 to improve gene alteration. We show that by fusing an ssODN template to the trans-activating RNA (tracrRNA), we recover precise genetic alterations, with increased integration and precision in vitro and in vivo. Finally, we show that this technology can be used to enhance gene conversion with other gene editing tools such as transcription activator like effector nucleases.

Genome-wide analysis of in vivo CcpA binding with and without its key co-factor HPr in the major human pathogen group A Streptococcus.

Catabolite control protein A (CcpA) is a master regulator of carbon source utilization and contributes to the virulence of numerous medically important Gram-positive bacteria. Most functional assessments of CcpA, including interaction with its key co-factor HPr, have been performed in nonpathogenic bacteria. In this study we aimed to identify the in vivo DNA binding profile of CcpA and assess the extent to which HPr is required for CcpA-mediated regulation and DNA binding in the major human pathogen group A Streptococcus (GAS). Using a combination RNAseq/ChIP-seq approach, we found that CcpA affects transcript levels of 514 of 1667 GAS genes (31%) whereas direct DNA binding was identified for 105 GAS genes. Three of the directly regulated genes encode the key GAS virulence factors Streptolysin S, PrtS (IL-8 degrading proteinase), and SpeB (cysteine protease). Mutating CcpA Val301 to Ala (strain 2221-CcpA-V301A) abolished interaction between CcpA and HPr and impacted the transcript levels of 205 genes (40%) in the total CcpA regulon. By ChIP-seq analysis, CcpAV301A bound to DNA from 74% of genes bound by wild-type CcpA, but generally with lower affinity. These data delineate the direct CcpA regulon and clarify the HPr-dependent and independent activities of CcpA in a key pathogenic bacterium.

Co-occurrence Interaction Networks of Extremophile Species Living in a Copper Mining Tailing.

Copper mining tailings are characterized by high concentrations of heavy metals and an acidic pH, conditions that require an extreme adaptation for any organism. Currently, several bacterial species have been isolated and characterized from mining environments; however, very little is known about the structure of microbial communities and how their members interact with each other under the extreme conditions where they live. This work generates a co-occurrence network, representing the bacterial soil community from the Cauquenes copper tailing, which is the largest copper waste deposit worldwide. A representative sampling of six zones from the Cauquenes tailing was carried out to determine pH, heavy metal concentration, total DNA extraction, and subsequent assignment of Operational Taxonomic Units (OTUs). According to the elemental concentrations and pH, the six zones could be grouped into two sectors: (1) the "new tailing," characterized by neutral pH and low concentration of elements, and (2) the "old tailing," having extremely low pH (~3.5) and a high concentration of heavy metals (mainly copper). Even though the abundance and diversity of species were low in both sectors, the Pseudomonadaceae and Flavobacteriaceae families were over-represented. Additionally, the OTU identifications allowed us to identify a series of bacterial species with diverse biotechnological potentials, such as copper bioleaching and drought stress alleviation in plants. Using the OTU information as a template, we generated co-occurrence networks for the old and new tailings. The resulting models revealed a rearrangement between the interactions of members living in the old and new tailings, and highlighted conserved bacterial drivers as key nodes, with positive interactions in the network of the old tailings, compared to the new tailings. These results provide insights into the structure of the soil bacterial communities growing under extreme environmental conditions in mines.

Deploying MMEJ using MENdel in precision gene editing applications for gene therapy and functional genomics.

Gene-editing experiments commonly elicit the error-prone non-homologous end joining for DNA double-strand break (DSB) repair. Microhomology-mediated end joining (MMEJ) can generate more predictable outcomes for functional genomic and somatic therapeutic applications. We compared three DSB repair prediction algorithms - MENTHU, inDelphi, and Lindel - in identifying MMEJ-repaired, homogeneous genotypes (PreMAs) in an independent dataset of 5,885 distinct Cas9-mediated mouse embryonic stem cell DSB repair events. MENTHU correctly identified 46% of all PreMAs available, a ∼2- and ∼60-fold sensitivity increase compared to inDelphi and Lindel, respectively. In contrast, only Lindel correctly predicted predominant single-base insertions. We report the new algorithm MENdel, a combination of MENTHU and Lindel, that achieves the most predictive coverage of homogeneous out-of-frame mutations in this large dataset. We then estimated the frequency of Cas9-targetable homogeneous frameshift-inducing DSBs in vertebrate coding regions for gene discovery using MENdel. 47 out of 54 genes (87%) contained at least one early frameshift-inducing DSB and 49 out of 54 (91%) did so when also considering Cas12a-mediated deletions. We suggest that the use of MENdel helps researchers use MMEJ at scale for reverse genetics screenings and with sufficient intra-gene density rates to be viable for nearly all loss-of-function based gene editing therapeutic applications.

Developing Strategies for Sustainable Medical Equipment Maintenance in Under-Resourced Settings.

Engineering technology plays a pivotal role in the delivery of health care in under-resourced countries by providing an infrastructure to improve patient outcomes. However, sustainability of these technologies is difficult in these settings oftentimes due to limited resources or training. The framework presented in this editorial focuses on establishing medical and laboratory equipment sustainability in developing countries and is comprised of four steps: 1) establishing reliable in-country relationships with stakeholders, 2) identifying needs for sustainable solutions locally, 3) exploring potential solutions and assessing their effort-to-impact ratios, and 4) working with strategic partners to implement solutions with clear performance metrics. By focusing on the sustainability of donated equipment instead of the equipment itself, this method presented distinguishes itself from other philanthropic endeavors in the field by seeking to establish preventive maintenance habits that can impact clinical outcomes of a community long term. Application of this methodology is reported in the Original Research Article "A Low-Cost Humidity Control System to Protect Microscopes in a Tropical Climate" by Asp et. al.

A Low-Cost Humidity Control System to Protect Microscopes in a Tropical Climate.

Introduction: A clean and functional microscope is necessary for accurate diagnosis of infectious diseases. In tropical climates, high humidity levels and improper storage conditions allow for the accumulation of debris and fungus on the optical components of diagnostic equipment, such as microscopes. Objective: Our objective was to develop and implement a low-cost, sustainable, easy to manage, low-maintenance, passive humidity control chamber to both reduce debris accumulation and microbial growth onto the optical components of microscopes. Methods: Constructed from easily-sourced and locally available materials, the cost of each humidity control chamber is approximately $2.35 USD. Relative humidity levels were recorded every 30 minutes over a period of 10 weeks from two chambers deployed at the Belize Vector and Ecology Center and the University of Belize. Results: The humidity control chamber deployed at the University of Belize maintained internal relative humidity at an average of 35.3% (SD = 4.2%) over 10 weeks, while the average external relative humidity was 86.4% (SD = 12.4%). The humidity control chamber deployed at the Belize Vector and Ecology Center effectively maintained internal relative humidity to an average of 54.5% (SD = 9.4%) over 10 weeks, while the average external relative humidity was 86.9% (SD = 12.9%). Conclusions: Control of relative humidity is paramount for the sustainability of medical equipment in tropical climates. The humidity control chambers reduced relative humidity to levels that were not conducive for fungal growth while reducing microscope contamination from external sources. This will likely extend the service life of the microscopes while taking advantage of low-cost, locally sourced components.

The Gene Sculpt Suite: a set of tools for genome editing.

The discovery and development of DNA-editing nucleases (Zinc Finger Nucleases, TALENs, CRISPR/Cas systems) has given scientists the ability to precisely engineer or edit genomes as never before. Several different platforms, protocols and vectors for precision genome editing are now available, leading to the development of supporting web-based software. Here we present the Gene Sculpt Suite (GSS), which comprises three tools: (i) GTagHD, which automatically designs and generates oligonucleotides for use with the GeneWeld knock-in protocol; (ii) MEDJED, a machine learning method, which predicts the extent to which a double-stranded DNA break site will utilize the microhomology-mediated repair pathway; and (iii) MENTHU, a tool for identifying genomic locations likely to give rise to a single predominant microhomology-mediated end joining allele (PreMA) repair outcome. All tools in the GSS are freely available for download under the GPL v3.0 license and can be run locally on Windows, Mac and Linux systems capable of running R and/or Docker. The GSS is also freely available online at www.genesculpt.org.

Robust activation of microhomology-mediated end joining for precision gene editing applications.

One key problem in precision genome editing is the unpredictable plurality of sequence outcomes at the site of targeted DNA double stranded breaks (DSBs). This is due to the typical activation of the versatile Non-homologous End Joining (NHEJ) pathway. Such unpredictability limits the utility of somatic gene editing for applications including gene therapy and functional genomics. For germline editing work, the accurate reproduction of the identical alleles using NHEJ is a labor intensive process. In this study, we propose Microhomology-mediated End Joining (MMEJ) as a viable solution for improving somatic sequence homogeneity in vivo, capable of generating a single predictable allele at high rates (56% ~ 86% of the entire mutant allele pool). Using a combined dataset from zebrafish (Danio rerio) in vivo and human HeLa cell in vitro, we identified specific contextual sequence determinants surrounding genomic DSBs for robust MMEJ pathway activation. We then applied our observation to prospectively design MMEJ-inducing sgRNAs against a variety of proof-of-principle genes and demonstrated high levels of mutant allele homogeneity. MMEJ-based DNA repair at these target loci successfully generated F0 mutant zebrafish embryos and larvae that faithfully recapitulated previously reported, recessive, loss-of-function phenotypes. We also tested the generalizability of our approach in cultured human cells. Finally, we provide a novel algorithm, MENTHU (http://genesculpt.org/menthu/), for improved and facile prediction of candidate MMEJ loci. We believe that this MMEJ-centric approach will have a broader impact on genome engineering and its applications. For example, whereas somatic mosaicism hinders efficient recreation of knockout mutant allele at base pair resolution via the standard NHEJ-based approach, we demonstrate that F0 founders transmitted the identical MMEJ allele of interest at high rates. Most importantly, the ability to directly dictate the reading frame of an endogenous target will have important implications for gene therapy applications in human genetic diseases.

Fishing for understanding: Unlocking the zebrafish gene editor's toolbox.

The rapid growth of the field of gene editing can largely be attributed to the discovery and optimization of designer endonucleases. These include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regular interspersed short palindromic repeat (CRISPR) systems including Cas9, Cas12a, and structure-guided nucleases. Zebrafish (Danio rerio) have proven to be a powerful model system for genome engineering testing and applications due to their external development, high fecundity, and ease of housing. As the zebrafish gene editing toolkit continues to grow, it is becoming increasingly important to understand when and how to utilize which of these technologies for maximum efficacy in a particular project. While CRISPR-Cas9 has brought broad attention to the field of genome engineering in recent years, designer endonucleases have been utilized in genome engineering for more than two decades. This chapter provides a brief overview of designer endonuclease and other gene editing technologies in zebrafish as well as some of their known functional benefits and limitations depending on specific project goals. Finally, selected prospects for additional gene editing tools are presented, promising additional options for directed genomic programming of this versatile animal model system.

Precision gene editing technology and applications in nephrology.

The expanding field of precision gene editing is empowering researchers to directly modify DNA. Gene editing is made possible using synonymous technologies: a DNA-binding platform to molecularly locate user-selected genomic sequences and an associated biochemical activity that serves as a functional editor. The advent of accessible DNA-targeting molecular systems, such as zinc-finger nucleases, transcription activator-like effectors (TALEs) and CRISPR-Cas9 gene editing systems, has unlocked the ability to target nearly any DNA sequence with nucleotide-level precision. Progress has also been made in harnessing endogenous DNA repair machineries, such as non-homologous end joining, homology-directed repair and microhomology-mediated end joining, to functionally manipulate genetic sequences. As understanding of how DNA damage results in deletions, insertions and modifications increases, the genome becomes more predictably mutable. DNA-binding platforms such as TALEs and CRISPR can also be used to make locus-specific epigenetic changes and to transcriptionally enhance or suppress genes. Although many challenges remain, the application of precision gene editing technology in the field of nephrology has enabled the generation of new animal models of disease as well as advances in the development of novel therapeutic approaches such as gene therapy and xenotransplantation.

ssDNA and the Argonautes: The Quest for the Next Golden Editor.

Genome engineering has gone mainstream because of breakthroughs in defining and harnessing naturally occurring, customizable DNA recognition cursors (protein or RNA-guided). At present, most gene editing relies on these cursors to direct custom DNA endonucleases to a specific genomic sequence to induce a double-strand break. New tools for genome engineering are continuously being explored, and another advance in DNA targeting has recently been described. Argonaute isolated from Natronobacterium gregoryi (NgAgo) is an ssDNA-based cursor that thus far has no known limitations in sequence recognition, shows promise for high specificity, and for many applications may represent a potentially more accessible genome-editing system over prior tools as it requires only a single, 24-base, 5' phosphorylated ssDNA for DNA targeting. Genome engineering is in a remarkable moment of unprecedented growth with exponential reduction in costs reminiscent of Moore's law in electronics. Many questions remain with regard to Argonaute utility in specific systems, but there is no doubt that genome engineering is expanding into new and exciting areas from synthetic biology to gene therapy.

FusX: A Rapid One-Step Transcription Activator-Like Effector Assembly System for Genome Science.

Transcription activator-like effectors (TALEs) are extremely effective, single-molecule DNA-targeting molecular cursors used for locus-specific genome science applications, including high-precision molecular medicine and other genome engineering applications. TALEs are used in genome engineering for locus-specific DNA editing and imaging, as artificial transcriptional activators and repressors, and for targeted epigenetic modification. TALEs as nucleases (TALENs) are effective editing tools and offer high binding specificity and fewer sequence constraints toward the targeted genome than other custom nuclease systems. One bottleneck of broader TALE use is reagent accessibility. For example, one commonly deployed method uses a multitube, 5-day assembly protocol. Here we describe FusX, a streamlined Golden Gate TALE assembly system that (1) is backward compatible with popular TALE backbones, (2) is functionalized as a single-tube 3-day TALE assembly process, (3) requires only commonly used basic molecular biology reagents, and (4) is cost-effective. More than 100 TALEN pairs have been successfully assembled using FusX, and 27 pairs were quantitatively tested in zebrafish, with each showing high somatic and germline activity. Furthermore, this assembly system is flexible and is compatible with standard molecular biology laboratory tools, but can be scaled with automated laboratory support. To demonstrate, we use a highly accessible and commercially available liquid-handling robot to rapidly and accurately assemble TALEs using the FusX TALE toolkit. Together, the FusX system accelerates TALE-based genomic science applications from basic science screening work for functional genomics testing and molecular medicine applications.

TrkB gene therapy by adeno-associated virus enhances recovery after cervical spinal cord injury.

Unilateral cervical spinal cord hemisection at C2 (C2SH) interrupts descending bulbospinal inputs to phrenic motoneurons, paralyzing the diaphragm muscle. Recovery after C2SH is enhanced by brain derived neurotrophic factor (BDNF) signaling via the tropomyosin-related kinase subtype B (TrkB) receptor in phrenic motoneurons. The role for gene therapy using adeno-associated virus (AAV)-mediated delivery of TrkB to phrenic motoneurons is not known. The present study determined the therapeutic efficacy of intrapleural delivery of AAV7 encoding for full-length TrkB (AAV-TrkB) to phrenic motoneurons 3 days post-C2SH. Diaphragm EMG was recorded chronically in male rats (n=26) up to 21 days post-C2SH. Absent ipsilateral diaphragm EMG activity was verified 3 days post-C2SH. A greater proportion of animals displayed recovery of ipsilateral diaphragm EMG activity during eupnea by 14 and 21 days post-SH after AAV-TrkB (10/15) compared to AAV-GFP treatment (2/11; p=0.031). Diaphragm EMG amplitude increased over time post-C2SH (p<0.001), and by 14 days post-C2SH, AAV-TrkB treated animals displaying recovery achieved 48% of the pre-injury values compared to 27% in AAV-GFP treated animals. Phrenic motoneuron mRNA expression of glutamatergic AMPA and NMDA receptors revealed a significant, positive correlation (r(2)=0.82), with increased motoneuron NMDA expression evident in animals treated with AAV-TrkB and that displayed recovery after C2SH. Overall, gene therapy using intrapleural delivery of AAV-TrkB to phrenic motoneurons is sufficient to promote recovery of diaphragm activity, adding a novel potential intervention that can be administered after upper cervical spinal cord injury to improve impaired respiratory function.

[Assessment of clinical competence in a pediatric residency with the Mini-Clinical Evaluation Exercise (Mini-CEX)]. / Evaluación de las competencias clínicas en una residencia de pediatría con el Mini-CEX (Mini-Clinical Evaluation Exercise).

PURPOSE: Assess the clinical competence of pediatric residents with the implementation of Mini-Clinical Evaluation Exercise (Mini-CEX), determining its validity, reliability, feasibility and satisfaction of examiners and residents. METHODS: 14 examiners and 8 residents of pediatrics took part. The Mini-CEX, a method based on direct observation of residents during their daily training, was used. A nine-point rating scale was used in order to evaluate their skills regarding medical interviewing, physical examination, professionalism, clinical judgment, counselling, organization, overall competence and satisfaction with the method. RESULTS: 181 observations were made, an average of 12.92 observations per examiner (range-2-39). Each examiner assessed 5.78 residents, (range 2-8). There was an average of 22.6 assessments per resident, range (18-30). The observations took place in outpatient clinic 38.7%, pediatric inpatient unit 19.3%, neonatal intensive care unit 17.1%, neonatal reception unit 14.4% and rooming-in 10.5%. The mean scores were: professionalism 7.15; interviewing 6.64; physical examination 6.67; clinical judgment 6.70; counselling 6.79 and organization 6.73. The overall competence score varied according to experience levels. Mean scores were: first-year residents 6.57; second-year residents 6.87 and third-year residents 7.3; p= 0.004. The score related to examiners's satisfaction was 7.89 and that of the residents was 7.74. The duration of the observation period was 28.35 minutes. Cronbach alfa coefficient was 0.97 showing the high reliability of the assessment method. The ANOVA score for overall competence of all examiners showed statistically significant differences, p <0.0001 in relation to stricter or more lenient judgment to evaluate skills. CONCLUSIONS: The implementation of the Mini-CEX in the Pediatrics Residency was feasible and positively accepted by residents and examiners. It allowed the assessment of different levels of performance among residents according to their experience, in every clinical setting of a pediatrician's practice. The variability criteria among examiners and the lack of constructive criticism are matters to be dealt with in future investigations.

Evaluación de las competencias clínicas en una residencia de pediatría con el Mini-CEX (Mini-Clinical Evaluation Exercise) / Assessment of clinical competence in a pediatric residency with the Mini-Clinical Evaluation Exercise (Mini-CEX)

Objetivo. Evaluar las competencias clínicas de los residentes de pediatría con la implementación del Mini-CEX, determinando su validez, confabilidad, factibilidad y la satisfacción de docentes y de residentes. Métodos. Participaron 14 docentes y 8 residentes. Se utilizó el Mini-CEX, método basado en la observación directa del desempeño del residente durante su práctica diaria, por parte de un docente. Resultados. Se realizaron 181 observaciones, media de 12,92 observaciones por cada docente. Cada docente evaluó a 5,78 residentes. Hubo una media de 22,6 evaluaciones por residente. Las observaciones se realizaron en consultorios externos 38,7%, internación pediátrica 19,3%, neonatología 17,1%, sala de recepción del recién nacido 14,4% y en internación conjunta 10,5%. Los puntajes promedios fueron: profesionalismo 7,15; entrevista 6,64; examen clínico 6,67; criterio clínico 6,70; asesoramiento 6,79 y organización 6,73. Los puntajes de competencia global variaron de acuerdo a los años de experiencia. Primer año 6,57; segundo 6,87 y tercero 7,3; p= 0,004. El puntaje de satisfacción de los docentes fue 7,89 y de los residentes 7,74. El tiempo de duración de las observaciones fue de 28,35 minutos. El coeficiente alfa de Cronbach fue de 0,97 lo que indica elevada confabilidad del método de evaluación. El ANOVA de puntajes de competencia global de todos los docentes mostró diferencias estadísticamente signifcativas, p <0,0001. Conclusiones. La implementación del Mini-CEX fue factible, bien aceptada por residentes y docentes, permitió valorar los diferentes niveles de desempeño de los residentes.

Purpose. Assess the clinical competence of pediatric residents with the implementation of Mini-Clinical Evaluation Exercise (Mini-CEX), determining its validity, reliability, feasibility and satisfaction of examiners and residents. Methods. 14 examiners and 8 residents of pediatrics took part. The Mini-CEX, a method based on direct observation of residents during their daily training, was used. A nine-point rating scale was used in order to evaluate their skills regarding medical interviewing, physical examination, professionalism, clinical judgment, counselling, organization, overall competence and satisfaction with the method. Results. 181 observations were made, an average of 12.92 observations per examiner (range-2-39). Each examiner assessed 5.78 residents, (range 2-8). There was an average of 22.6 assessments per resident, range (18-30). The observations took place in outpatient clinic 38.7%, pediatric inpatient unit 19.3%, neonatal intensive care unit 17.1%, neonatal reception unit 14.4% and rooming-in 10.5%. The mean scores were: professionalism 7.15; interviewing 6.64; physical examination 6.67; clinical judgment 6.70; counselling 6.79 and organization 6.73. The overall competence score varied according to experience levels. Mean scores were: frst-year residents 6.57; second-year residents 6.87 and third-year residents 7.3; p= 0.004. The score related to ex-aminers's satisfaction was 7.89 and that of the residents was 7.74. The duration of the observation period was 28.35 minutes. Cronbach alfa coeffcient was 0.97 showing the high reliability of the assessment method. The ANOVA score for overall competence of all examiners showed statistically signifcant differences, p <0.0001 in relation to stricter or more lenient judgment to evaluate skills. Conclusions. The implementation of the Mini-CEX in the Pediatrics Residency was feasible and positively accepted by residents and examiners. It allowed the assessment of different levels of performance among residents according to their experience, in every clinical setting of a pediatrician's practice. The variability criteria among examiners and the lack of constructive criticism are matters to be dealt with in future investigations.

Escherichia coli ehl1 gene-positive serotype O18ac:H31 associated with an outbreak of diarrhea in a neonatal nursery in Neuquén City, Argentina.

Between 9 October and 12 November 1996, an outbreak of bloody diarrhea occurred in the neonatal nursery ward of the Policlínico Neuquén, in Neuquén, a city in the southwestern region of Argentina. Seven patients of the intermediate care unit were affected. Isolates of Escherichia coli O18ac:H31 that were non-lactose and -sorbitol fermenting were recovered from outbreak cases. Although the strains were negative for a number of virulence factors typically found in diarrheagenic groups of E. coli, all isolates from the present neonatal outbreak possessed the enterohemolysin gene, ehl1. All isolates showed resistance to the antibiotics ampicillin and chloramphenicol. These isolates showed a low adherence property in HeLa cells without any recognizable pattern. In order to evaluate the outbreak dissemination in the neonatology ward, a prevalence study was conducted on 13 November. Stool specimens were obtained from 16 neonates hospitalized in the sector and from 33 medical staff members. E. coli isolates with identical biochemical characteristics of the outbreak strain were recovered from 11 of 16 inpatients and from 4 of 33 staff members during the prevalence study. A total of 15 E. coli strains recovered both from the outbreak and the prevalence study were processed by random amplified polymorphic DNA (RAPD)-PCR and pulsed-field gel electrophoresis (PFGE). By RAPD-PCR 14 of 15 strains showed patterns with 85 to 100% similarity, and by PFGE these strains were identical, each showing a unique pattern with 15 bands between 40 and 400 kb. One strain isolated from a nurse during the prevalence study presented a pattern not related to the others, and this was characterized as E. coli O81:HNM resistant to ampicillin only and negative for all the virulence factors studied. This outbreak occurred despite strict regulations in place to prevent cross-infection in the hospital. Postoutbreak prevalence studies were performed weekly thereafter without detecting new cases.

Estudio descriptivo de la población atendida en consultorios externos de pediatría / Descriptive study of the population attendedd in pediatric external consulting rooms

Introducción.La atención del consultorio externo de pediatría es la actividad principal de la mayoría de los pediatras.No obstante,en las residencias pediátricas se pone mayor énfasis en la formación de internistas.Para adecuar los contenidos del programa formativo de la residencia y optimizar la rotación por consultorios externos se realizó el presente estudio.Objetivos.Conocer las características de la población atendida en consultorios externos de pediatría,los motivos de consulta y las patologías prevalentes.Conocer el número de pacietnes en control del niño sano y de consultass por demanda espontánea,así como su distribución por edad y sexo.Estimar el número de estudios complementarios de pacientes internados y de pacientes medicados con antibióticos,Comparar los resultados de atención de los médicos de planta t de los residentes y evaluar la labor de éstos.Pacientes y métodos.Estudio descriptivo de los pacientes atendidos en consultorios externos del servicio de pediatría y neonatología desde el 01/04/99 al 31/03/00.se incluyeron todos los pacientes menores de 16 años que consultaron para control del niño sao por demanda espontánea.Los niños fueron atendidos por pediatras del plantel o por médicos residentes.Se confeccionó una planilla para el registro de datos de los pacientes atendidis por cada médico y se trasladaron a la base de datos en Access.Conclusiones.el 75 por ciento de los pacientes atendidos fueron menores de 6 años y el CNS se mantuvo estable,pero exitió una marcada variabilidad estacional en el número de consultas por demanda espontánea.Las principales causas de internación fueron los accidentes,gastroenteritis,fiebre sin fici,síndrome bronquiolar y abdomen agudo.Los médicos residentes atendieron un número significativo de niños.Dedicaron el 50 por ciento de su tiempo a la atención del consultorio.Pensamos que su rotación por consultorios externos ofrece oportunidades suficientes de capacitación,No existieron diferencias estadísticamente significativas entre el número de estudios complementarios solicitados por residentes y mñedicos de planta.En cambio,se observaron diferencia en cuanto a la indicación de antibióticos y al porcentaje de pacientes internados

Estudio descriptivo de la población atendida en consultorios externos de pediatría / Descriptive study of the population attendedd in pediatric external consulting rooms

Introducción.La atención del consultorio externo de pediatría es la actividad principal de la mayoría de los pediatras.No obstante,en las residencias pediátricas se pone mayor énfasis en la formación de internistas.Para adecuar los contenidos del programa formativo de la residencia y optimizar la rotación por consultorios externos se realizó el presente estudio.Objetivos.Conocer las características de la población atendida en consultorios externos de pediatría,los motivos de consulta y las patologías prevalentes.Conocer el número de pacietnes en control del niño sano y de consultass por demanda espontánea,así como su distribución por edad y sexo.Estimar el número de estudios complementarios de pacientes internados y de pacientes medicados con antibióticos,Comparar los resultados de atención de los médicos de planta t de los residentes y evaluar la labor de éstos.Pacientes y métodos.Estudio descriptivo de los pacientes atendidos en consultorios externos del servicio de pediatría y neonatología desde el 01/04/99 al 31/03/00.se incluyeron todos los pacientes menores de 16 años que consultaron para control del niño sao por demanda espontánea.Los niños fueron atendidos por pediatras del plantel o por médicos residentes.Se confeccionó una planilla para el registro de datos de los pacientes atendidis por cada médico y se trasladaron a la base de datos en Access.Conclusiones.el 75 por ciento de los pacientes atendidos fueron menores de 6 años y el CNS se mantuvo estable,pero exitió una marcada variabilidad estacional en el número de consultas por demanda espontánea.Las principales causas de internación fueron los accidentes,gastroenteritis,fiebre sin fici,síndrome bronquiolar y abdomen agudo.Los médicos residentes atendieron un número significativo de niños.Dedicaron el 50 por ciento de su tiempo a la atención del consultorio.Pensamos que su rotación por consultorios externos ofrece oportunidades suficientes de capacitación,No existieron diferencias estadísticamente significativas entre el número de estudios complementarios solicitados por residentes y mñedicos de planta.En cambio,se observaron diferencia en cuanto a la indicación de antibióticos y al porcentaje de pacientes internados

Relación entre el receptor soluble de unterleucina-2 y sepsis neonatal / Relationship between soluble interlukin 2 receptor and the newborn sepsis

Introducción.Diversas citocina actúan como mediadoras de la respuesta inmune específica.El recepto solublr de interleucina -2(RSIL-2)es considerado un marcador de la activación del sistema inmuney el aumento de su concentración en un neonato con sospecha de sepsis podría apoyar el diagnóstico.Objetivo.determinar la relación entre el RSIL-2 y la sepsis neonatal y determinar si la elevación del nivel serico de RSIL-2 tiene valor para el diagnóstico de sepsis.Conclusiones.Se encontró una asociación entre la elevacióndel RSIL-2 y la sepsis neonatal.La determinación entre la elevación del RSIL-2 y la sepsis neonatal.La determinación del RSIL-2 en la evaluación inicial de un paciente por sospecha de sepsis puede contribuir con el diagnóstico de sepsis neonatal