Study protocol to investigate biomolecular muscle profile as predictors of long-term urinary incontinence in women with gestational diabetes mellitus.

BACKGROUND: Pelvic floor muscles (PFM) and rectus abdominis muscles (RAM) of pregnant diabetic rats exhibit atrophy, co-localization of fast and slow fibers and an increased collagen type I/III ratio. However, the role of similar PFM or RAM hyperglycemic-related myopathy in women with gestational diabetes mellitus (GDM) remains poorly investigated. This study aims to assess the frequency of pelvic floor muscle disorders and pregnancy-specific urinary incontinence (PS-UI) 12 months after the Cesarean (C) section in women with GDM. Specifically, differences in PFM/RAM hyperglycemic myopathy will be evaluated. METHODS: The Diamater is an ongoing cohort study of four groups of 59 pregnant women each from the Perinatal Diabetes Research Centre (PDRC), Botucatu Medical School (FMB)-UNESP (São Paulo State University), Brazil. Diagnosis of GDM and PS-UI will be made at 24-26 weeks, with a follow-up at 34-38 weeks of gestation. Inclusion in the study will occur at the time of C-section, and patients will be followed at 24-48 h, 6 weeks and 6 and 12 months postpartum. Study groups will be classified as (1) GDM plus PS-UI; (2) GDM without PS-UI; (3) Non-GDM plus PS-UI; and (4) Non-GDM without PS-UI. We will analyze relationships between GDM, PS-UI and hyperglycemic myopathy at 12 months after C-section. The mediator variables to be evaluated include digital palpation, vaginal squeeze pressure, 3D pelvic floor ultrasound, and 3D RAM ultrasound. RAM samples obtained during C-section will be analyzed for ex-vivo contractility, morphological, molecular and OMICS profiles to further characterize the hyperglycemic myopathy. Additional variables to be evaluated include maternal age, socioeconomic status, educational level, ethnicity, body mass index, weight gain during pregnancy, quality of glycemic control and insulin therapy. DISCUSSION: To our knowledge, this will be the first study to provide data on the prevalence of PS-UI and RAM and PFM physical and biomolecular muscle profiles after C-section in mothers with GDM. The longitudinal design allows for the assessment of cause-effect relationships between GDM, PS-UI, and PFMs and RAMs myopathy. The findings may reveal previously undetermined consequences of GDM.

Adipocyte proteome and secretome influence inflammatory and hormone pathways in glioma.

Gliomas represent the most common primary malignant brain tumors in adults, with an extremely poor prognosis. Among several risk factors, lifestyle was also recently identified as a major risk factor for the development of primary glioma. In the present study, we explore the relationship between obesity and glioma in a cellular model. Thus, we have study the influence of adipocytes secretome on glioma cell line GL261. Using the 3T3-L1 adipocyte cell line, and its conditioned medium (adipokines-enriched medium), we showed that adipocyte-released factors relate with glioma angiogenic, growth, hormones and metabolic behavior by MALDI-TOF-MS and proteomic array analysis. In a first view, STI1, hnRNPs and PGK1 are under expressed on CGl. Similarly, both carbonic anhydrase and aldose reductase are even suppressed in glioma cells that grown under adipokines-enriched environment. Contrariwise, RFC1, KIF5C, ANXA2, N-RAP and RACK1 are overexpressed in GL261 cell the in the presence of the adipokines-enriched medium. We further identified the factors that are released by adipocyte cells, and revealed that several pro-inflammatory and angiogenic factors, such as IL-6, IL-11, LIF, PAI-1, TNF-α, endocan, HGF, VEGF IGF-I, were secreted to the medium into a high extent, whereas TIMP-1 and SerpinE1 were under expressed on CGl. This study discloses an interesting in vitro model for the study of glioma biology under a "obesity" environment, that can be explored for the understanding of cancer cells biology, for the search of biomarkers, prognostic markers and therapeutic approaches.

Simulador de bajo costo para el entrenamiento en la colocación de accesos vasculares periféricos (AVP) en pediatría / Low-cost simulator for the training of peripheral venous access (PVA) placement in pediatrics

Introducción: El entrenamiento en simuladores permite aprender procedimientos en un marco controlado que protege la seguridad de los pacientes pediátricos y que se integra como una instancia de aprendizaje previa a la realidad con el paciente. En el Centro de Simulación del Hospital de Pediatría Juan P. Garrahan se diseñan y validan simuladores de bajo costo artesanales que permiten contar con recursos propios y de reposición inmediata. Objetivo: Describir el diseño de un simulador de bajo costo para la colocación de accesos vasculares periféricos y reportar la experiencia inicial de validación. Métodos: Se diseñó un simulador artesanal inanimado de punción venosa periférica (parte de miembro superior de un niño de edad aproximada de 8 años) con materiales de bajo costo. En una segunda fase y con el propósito de validarlo, se solicitó la colocación del acceso vascular -en el modelo simulado- a enfermeros del hospital que trabajan en el área de internación (usuarios expertos). Al final de cada experiencia, cada operador reportó su opinión sobre el realismo del modelo y utilidad de la experiencia en forma anónima. Resultados Participaron 43 enfermeros; entre el 75 a 90% de las veces, las repuestas fueron "se parece mucho" o "es igual" a la experiencia real de colocación de un acceso venoso a un niño. El 85% expresó que esta práctica permitía mejorar la destreza, y el 100% que el modelo puede ayudar a enseñarla en forma efectiva. Conclusiones El desarrollo de modelos de simuladores de bajo costo para usos específicos, de baja tecnología, resulta importante para el entrenamiento de habilidades. La aceptación por parte de los usuarios calificados y expertos fue muy buena, encontrando en un alto porcentaje similitud con la realidad (AU)

Introduction: Training on simulators allows for the learning of procedures within a controlled framework that protects the safety of pediatric patients and provides a learning moment previous to working with a real patient. In the Simulation Center at Hospital de Pediatría Juan P. Garrahan low-cost simulators are designed and validated that allow for proper resources and immediate replacement. Aim: To describe the design of a low-cost simulator for the placement of peripheral vascular Access and to report the initial experience with the validation of the device. Methods: An inanimate simulator for peripheral puncture (the upper limb of a child of approximately 8 years of age) was designed using lowcost materials. In a second phase with the purpose of validating the device, nurses of the hospital working in the inpatient area (expert users) were asked to insert a venous catheter in the simulation model. At the end of each procedure, each operator was asked to anonymously give their opinion. Results: 43 nurses participated; between 75 and 90% of times, the answers were "it is very similar "or "it is the same as the experience of placing a real venous catheter in a child. Overall, 85% felt that the training improved their skills and 100% considered that the model may be effective in the teaching process. Conclusions: The development of low-cost, low-technology simulation models is important in the training of skills. Acceptance by qualified users and experts was very good. A high degree of similarity with reality was reported (AU)

Stress enhances the sensitivity of Salmonella enterica serovar Typhimurium to bacteriocins.

AIMS: The aim of this study was to evaluate the potential application of bacteriocins against Gram-negative bacteria when associated with others food preservation methods. METHODS AND RESULTS: Salmonella was subjected to heat, cold, acid and chemical (with ethylenediaminetetracetate and trisodium phosphate) stresses. Then, the cells were recovered and subjected to treatment with bacteriocins (500 AU ml(-1) ) for 6 h. Heat and cold stress were those that promoted more sensitization to bactericidal activity of nisin. Under the same conditions, bovicin HC5 acted more rapidly than nisin reducing the number of viable cells to undetectable levels after 20 min of treatment. Similar results with use of nisin only were observed after 6 h of treatment. CONCLUSIONS: Stress conditions used in food industry, such as temperature and pH, and use of chelating agents or membrane disruptors, sensitized Salmonella Typhimurium cells to bacteriocins produced by lactic acid bacteria, such as nisin and bovicin HC5. SIGNIFICANCE AND IMPACT OF THE STUDY: Food preservation methods sensitized Gram-negative bacteria to bacteriocins activity, which demonstrate the potential of nisin and bovicin HC5 to inhibit the growth of Salmonella.

Differences in the antibacterial activity of nisin and bovicin HC5 against Salmonella Typhimurium under different temperature and pH conditions.

AIMS: To compare the action of nisin and bovicin HC5 in combination with EDTA on Salmonella Typhimurium under different environmental conditions. METHODS AND RESULTS: Salmonella Typhimurium was treated in BHI broth containing EDTA (1·5 mmol l(-1)) and nisin or bovicin HC5 (200 AU ml(-1)) under different pH and temperature conditions, and according to a central composite design with two factors (temperature and pH). Cell viability was evaluated on plate count agar for 48 h. The combination of nisin or bovicin HC5 with EDTA was able to inhibit the growth of Salmonella, but the temperature and pH conditions promoting inhibition were distinct for each bacteriocin. Nisin was bactericidal over a broad range of temperature and pH, while bovicin HC5 was bacteriostatic in most conditions and bactericidal only in specific conditions (pH >6·0 and temperature >30°C). Salmonella Typhimurium did not show tolerance to bovicin HC5 or cross-tolerance between these lantibiotics. CONCLUSIONS: Nisin and bovicin HC5 both inhibited the growth of Salmonella, but the activity of each bacteriocin was differently influenced by environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Nisin and bovicin HC5 have the potential to inhibit the growth of Salmonella, but environmental conditions should be considered to establish optimal conditions for its application.

Seguridad en el traslado intrahospitalario de pacientes críticos en pediatría / Safety of interhospital transfers of critical care patients in pediatrics

La unidad de cuidados intensivos (UCI) es el ámbito más seguro para la atención de pacientes críticamente enfermos. Sin embargo, hay situaciones en las que el paciente debe ser trasladado a algún otro lugar del hospital, pudiendo incrementar el riesgo para eventos adversos. El objetivo es describir la implementación de un programa de capacitación y realizar un relevamiento de los traslados de pacientes de UCI. Estudio descriptivo y prospectivo. Se incluyeron todos los pacientes trasladados desde las UCI 44-45-72 y 65 durante el periodo de Julio 2012 a Junio 2013. Se diseñó una lista de chequeo, con datos de cada paciente, material para el traslado y registro de efectos adversos. Se agruparon por gravedad en Grupo I: Sin requerimientos de inotrópicos y sin asistencia respiratoria mecánica (ARM) y Grupo II: Requerimientos de ARM y/o inotrópicos (A Estables, B Inestable). Se realizó capacitación del personal encargado de los traslados para completar las listas de chequeo y recomendaciones sobre traslado seguro. Se documentaron 104 traslados y se distribuyeron en Grupo IA 32%, 2A 61% y 2B 11%. Se pesquisó un total de 61 (58.65%) eventos adversos. Se registraron eventos adversos en 58.65% de los traslados y solo 47.11% de los traslados fueron realizados en condiciones adecuadas (AU)

The intensive care unit (ICU) is the safest environment for the care of critically ill patients. Nevertheless, in certain settings the patients have to be transferred to other sectors of the hospital, which may increase the risk of adverse events. With the aim to describe the implementation of a training program and to assess the transfer of ICU patients a descriptive and prospective study was conducted. All patients transferred from ICUs 44-45,72, and 65 over the period July 2012 to June 2013 were included. A checklist was developed with patient data, materials included in the transfer, and recording of adverse events. Patients were categorized according to severity into Group I: No need for inotropics and mechanical ventilation (MV) and Group II: Need for MV and/or inotropics (A Stable, B Unstable). Personnel in charge of the transfers were trained in the filling out of the checklists and recommendations for safe transfers. One hundred and four transfers were registered; 32% of the patients were in Group IA, 61% in Group 2A, and 11% in 2B. A total of 61 (58.65%) adverse events were observed. Adverse events were registered in 58.65% of the transfers and only 47.11% of the transfers were performed under adequate conditions (AU)

Peptide mimicking antigenic and immunogenic epitope of neuwiedase from Bothrops neuwiedi snake venom.

Peptides derived from a phage display library may mimic essential features of epitopes (mimotopes), including their immunogenicity. A recombinant peptide library of 12 amino acids displayed on the phage capsid was used to obtain peptides that mimic epitopes of antigens that are reactive to specific polyclonal antibodies anti-neuwiedase (NEU), a toxin from Bothrops neuwiedi snake venom. These polyclonal antibodies are protective against NEU activity and were used as target for the peptide library biopannings, resulting in the selection of 80 peptides. Antibody-binding epitopes were obtained by sequence alignment with the primary and tertiary structures of the NEU protein. Antigenicity and specificity of the mimotopes mixture were confirmed by dot blot, immuno dot blot, plaque reduction and Western blot assays. Their immunogenicity was demonstrated by immunization of BALB/c mice and ELISA tests. The NEU toxin is an important antigen that has many common structural regions to several toxic venom metalloproteinases, in which two epitope regions have been detected. The two mapped epitopes were found in primary sequences of several snake venom toxins, thus demonstrating the potential application of these NEU mimotopes as possible antigen components that are toxicity free.

Rapid detection of efflux pumps and their relation with drug resistance in yeast cells.

BACKGROUND: Cell drug resistance can be due to the presence of active efflux pumps (AEP). Identification of yeast cells with a resistance phenotype is important either from a clinical, agricultural or biotechnological point of view. Rapid and reliable methods to detect AEP can be therefore very useful. METHODS: Some yeast cells change their staining by calcein-AM, BCECF-AM, rhodamine 123 and DiOC(5), when pretreated with verapamil, CCCP or ATP depletion, or when pretreated with specific antimicrobial agents. This fact may be interpreted as an indication of the presence/absence of AEP. Six yeast species were tested with a flow cytometric method (FCM) and an epifluorescence microscopic method (EFM), and ten other species were evaluated only by EFM. The minimum inhibitory concentration (MIC) of penconazol, benomyl and cycloheximide for Saccharomyces cerevisiae and Kluyveromyces marxianus, were determined by growth inhibition on solid medium and were compared to the staining changes detected by FCM. RESULTS: The FCM and the EFM allowed the detection of AEP in all the yeast species tested. High MIC values for a drug were related with the presence of at least one AEP indicated by the cytometric data. CONCLUSIONS: The FCM revealed to be a robust assay whereas the EFM can be used as a preliminary test. It is possible to identify resistance/sensitivity patterns in yeast cells through cytometric detection methods of different efflux pumping systems.

Flow cytometric assessment of cell structural and functional changes induced by acetic acid in the yeasts Zygosaccharomyces bailii and Saccharomyces cerevisiae.

Flow cytometry (FCM) was used with different viability dyes to assess changes in cell structure and function induced by acetic acid (AA) in populations of Zygosaccharomyces bailii (AA resistant) and Saccharomyces cerevisiae (AA sensitive). Kinetic changes in esterase activity, intracellular dye processing, and membrane integrity were monitored, and to detect those changes we used three assays involving fluorescein diacetate hydrolysis, FUN-1 processing, and propidium iodide exclusion, respectively. In S. cerevisiae, the decrease in the ability to process FUN-1 preceded the decrease in esterase activity, and there was loss of cell membrane integrity after incubation with AA. In Z. bailii, with higher AA concentrations, there was a similar decrease in the ability to process FUN-1, which also preceded the loss of cell membrane integrity. Changes in esterase activity in this yeast induced by AA treatment could not be monitored because the changes occurred independently of the presence of the acid. For control samples (untreated cells killed with 10% v/v of AA), the percentages of nonaltered cells as estimated by FCM and percentages of viable cells as estimated by colony forming unit (CFU) counts were identical. However, for cell samples treated for short periods with 3% (v/v) or less of AA, none of the dyes produced FCM results comparable to those produced by CFU counts.