PACAP binds conserved receptors and modulates cytokine gene expression and protein secretion in trout cell lines.

Antimicrobial peptides (AMPs) are an alternative to antibiotics for treatment and prevention of infections with a lower risk of bacterial resistance. Pituitary adenylate cyclase activating polypeptide (PACAP) is an outstanding AMP with versatile effects including antimicrobial activity and modulation of immune responses. The objective of this research was to study PACAP immunomodulatory effect on rainbow trout cell lines infected with Aeromonas salmonicida. PACAP from Clarias gariepinus (PACAP1) and a modified PACAP (PACAP5) were tested. RT-qPCR results showed that il1b and il8 expression in RTgutGC was significantly downregulated while tgfb expression was upregulated after PACAP treatment. Importantly, the concentration of IL-1ß and IFN-γ increased in the conditioned media of RTS11 cells incubated with PACAP1 and exposed to A. salmonicida. There was a poor correlation between gene expression and protein concentration, suggesting a stimulation of the translation of IL-1ß protein from previously accumulated transcripts or the cleavage of accumulated IL-1ß precursor. In-silico studies of PACAP-receptor interactions showed a turn of the peptide characteristic of PACAP-PAC1 interaction, correlated with the higher number of interactions observed with this specific receptor, which is also in agreement with the higher PACAP specificity described for PAC1 compared to VPAC1 and VPACA2. Finally, the in silico analysis revealed nine amino acids related to the PACAP receptor-associated functionality.

Innate and adaptive immune response of Rainbow trout (Oncorhynchus mykiss) naturally infected with Yersinia ruckeri.

Rainbow trout is an important fish species for Peruvian artisanal aquaculture, comprising over 60 % of the total aquaculture production. However, their industry has been highly affected by several bacterial agents such as Yersinia ruckeri. This pathogen is the causative agent of Enteric Redmouth Disease, and causes high mortality in fingerlings and chronic infection in adult rainbow trout. To date, the immune response of rainbow trout against Y. ruckeri has been well studied in laboratory-controlled infection studies (i.e. intraperitoneal infection, bath immersion), however, the immune response during natural infection has not been explored. To address this, in this study, 35 clinically healthy O. mykiss without evidence of lesions or changes in behavior and 32 rainbow trout naturally infected by Y. ruckeri, were collected from semi-intensive fish farms located in the Central Highlands of Peru. To evaluate the effect on the immune response, RT-qPCR, western blotting, and ELISA were conducted using head kidney, spleen, and skin tissues to evaluate the relative gene expression and protein levels. Our results show a significant increase in the expression of the pro-inflammatory cytokines il1b, tnfa, and il6, as well as ifng in all three tissues, as well as increases in IL-1ß and IFN-γ protein levels. The endogenous pathway of antigen presentation showed to play a key role in defense against Y. ruckeri, due to the upregulation of mhc-I, tapasin, and b2m transcripts, and the significant increase of Tapasin protein levels in infected rainbow trout. None of the genes associated with the exogenous pathway of antigen presentation showed a significant increase in infected fish, suggesting that this pathway is not involved in the response against this intracellular pathogen. Finally, the transcripts of immunoglobulins IgM and IgT did not show a modulation, nor were the protein levels evaluated in this study.

Role of cold shock proteins B and D in Aeromonas salmonicida subsp. salmonicida physiology and virulence in lumpfish (Cyclopterus lumpus).

Cold shock proteins (Csp) are pivotal nucleic acid binding proteins known for their crucial roles in the physiology and virulence of various bacterial pathogens affecting plant, insect, and mammalian hosts. However, their significance in bacterial pathogens of teleost fish remains unexplored. Aeromonas salmonicida subsp. salmonicida (hereafter A. salmonicida) is a psychrotrophic pathogen and the causative agent of furunculosis in marine and freshwater fish. Four csp genes (cspB, cspD, cspA, and cspC) have been identified in the genome of A. salmonicida J223 (wild type). Here, we evaluated the role of DNA binding proteins, CspB and CspD, in A. salmonicida physiology and virulence in lumpfish (Cyclopterus lumpus). A. salmonicida ΔcspB, ΔcspD, and the double ΔcspBΔcspD mutants were constructed and characterized. A. salmonicida ΔcspB and ΔcspBΔcspD mutants showed a faster growth at 28°C, and reduced virulence in lumpfish. A. salmonicida ΔcspD showed a slower growth at 28°C, biofilm formation, lower survival in low temperatures and freezing conditions (-20°C, 0°C, and 4°C), deficient in lipopolysaccharide synthesis, and low virulence in lumpfish. Additionally, ΔcspBΔcspD mutants showed less survival in the presence of bile compared to the wild type. Transcriptome analysis revealed that 200, 37, and 921 genes were differentially expressed in ΔcspB, ΔcspD, and ΔcspBΔcspD, respectively. In ΔcspB and ΔcspBΔcspD virulence genes in the chromosome and virulence plasmid were downregulated. Our analysis indicates that CspB and CspD mostly act as a transcriptional activator, influencing cell division (e.g., treB), virulence factors (e.g., aexT), and ultimately virulence.

The effects of Pediococcus acidilactici MA18/5M on growth performance, gut integrity, and immune response using in vitro and in vivo Pacific salmonid models.

Microbial management is central to aquaculture's efficiency. Pediococcus acidilactici MA18/5M has shown promising results promoting growth, modulation of the immune response, and disease resistance in many fishes. However, the mechanisms through which this strain confers health benefits in fish are poorly understood, particularly in Pacific salmonid models. Briefly, the aims of this study were to i) assess the protective effects of P. acidilactici MA18/5M by examining gut barrier function and the expression of tight junction (TJ) and immune genes in vitro and in vivo, and ii) to determine the protective effects of this strain against a common saltwater pathogen, Vibrio anguillarum J382. An in vitro model of the salmonid gut was employed utilizing the cell line RTgutGC. Barrier formation and integrity assessed by TEER measurements in RTgutGC, showed a significant decrease in resistance in cells exposed only to V. anguillarum J382 for 24 h, but pre-treatment with P. acidilactici MA18/5M for 48 h mitigated these effects. While P. acidilactici MA18/5M did not significantly upregulate tight junction and immune molecules, pre-treatment with this strain protected against pathogen-induced insults to the gut barrier. In particular, the expression of ocldn was significantly induced by V. anguillarum J382, suggesting that this molecule might play a role in the host response against this pathogen. To corroborate these observations in live fish, the effects of P. acidilactici MA18/5M was evaluated in Chinook salmon reared in real aquaculture conditions. Supplementation with P. acidilactici MA18/5M had no effect on Chinook salmon growth parameters after 10 weeks. Interestingly, histopathological results did not show alterations associated with P. acidilactici MA18/5M supplementation, indicating that this strain is safe to be used in the industry. Finally, the expression pattern of transcripts encoding TJ and immune genes in all the treatments suggest that variation in expression is more likely to be due to developmental processes rather than P. acidilactici MA18/5M supplementation. Overall, our results showed that P. acidilactici MA18/5M is a safe strain for use in fish production, however, to assess the effects on growth and immune response previously observed in other salmonid species, an assessment in adult fish is needed.

Consultas tardías en los servicios de emergencias pediátricas durante la pandemia por COVID-19 en Uruguay, entre el 13 de marzo y el 29 de julio de 2020

En diciembre de 2019, en Wuhan, China, se detectaron los primeros casos de SARS-CoV-2. En Uruguay, desde el 16 de marzo de 2020 se suspendieron las actividades de enseñanza, deportivas y espectáculos públicos. Varios países reportaron una marcada disminución de las visitas a urgencias. Algunos niños presentaron enfermedades ocasionales o descompensaciones de enfermedades crónicas, consultando en forma tardía con el riesgo que ello implica. El objetivo de este trabajo es realizar una descripción de las consultas tardías durante la pandemia. Se realizó un estudio multicéntrico y descriptivo entre el 13 de marzo y el 29 de julio de 2020. Se definió consulta tardía como los ingresos por injurias agudas con más de 6 horas de evolución, fiebre mayor a 72 horas de evolución, dificultad respiratoria con más de 12 horas de evolución, síntomas agudos, como dolor abdominal, de más de 24 horas de evolución, síntomas de más de 12 horas de evolución en niños con enfermedades crónicas que determinaron descompensación e ingreso. Se incluyeron 27 centros. Se registraron un total de 34.260 consultas en urgencia, se incluyeron 189 niños para el estudio. El promedio de edad fue de 6 años; 17 pacientes requirieron ingreso a unidad de cuidados intensivos (UCI). Predominó la apendicitis entre los diagnósticos al alta. Esta investigación puso en evidencia la existencia de consultas tardías en nuestro país. Esto contribuye a ponderar el impacto negativo de la pandemia en la población pediátrica.

Summary: In December 2019, the first cases of SARS-CoV-2 were detected in Wuhan. In Uruguay, since March 16, teaching, sports and public entertainment activities were suspended. Several countries reported a marked decrease in emergency room visits. Some children presented occasional illnesses or decompensations from chronic illnesses, consulting late with the risk that this implies. The objective of the work is to make a description of late consultations during the pandemic. A multicenter and descriptive study was carried out between March 13 and July 29, 2020. "Late consultation" was defined as admissions for: Acute injuries with more than 6 hours of evolution, fever greater than 72 hours of evolution, difficulty respiratory disease with more than 12 hours of evolution, acute symptoms such as abdominal pain of more than 24 hours of evolution, symptoms of more than 12 hours of evolution in children with chronic diseases that determined decompensation and admission. 27 centers were included. A total of 34260 emergency consultations were registered, 189 children were included for the study. The average age was 6 years. 17 patients required admission to the ICU. Appendicitis predominated among the diagnoses at discharge. This research revealed the existence of late consultations in our country. This helps to weigh the negative impact of the pandemic on the pediatric population.

Em dezembro de 2019, em Wuhan, foram detectados os primeiros casos de SARS-CoV-2. No Uruguai, desde 16 de março, as atividades de ensino, esporte e entretenimento público foram suspensas. Vários países relataram uma diminuição acentuada nas visitas ao pronto-socorro. Algumas crianças apresentavam doenças ocasionais ou descompensações de doenças crônicas, consultando tardiamente os riscos que isso implica. O objetivo do trabalho é fazer uma descrição das consultas tardias durante a pandemia. Um estudo multicêntrico e descritivo foi realizado entre 13 de março e 29 de julho de 2020. Consulta tardia foi definida como internações por: Lesões agudas com mais de 6 horas de evolução, febre maior que 72 horas de evolução, dificuldade respiratória com mais de 12 horas de evolução, sintomas agudos como dor abdominal com mais de 24 horas de evolução, sintomas com mais de 12 horas de evolução em crianças com doenças crônicas que determinaram descompensação e internação. 26 centros foram incluídos. Um total de 34.260 consultas de emergência foram registradas, 189 crianças foram incluídas no estudo. A idade média era de 6 anos. 17 pacientes necessitaram de internação na UTI. Apendicite predominou entre os diagnósticos na alta. Esta pesquisa revelou a existência de consultas tardias em nosso país. Isso ajuda a pesar o impacto negativo da pandemia na população pediátrica.

Comparative Genomic Analysis of Virulent Vibrio (Listonella) anguillarum Serotypes Revealed Genetic Diversity and Genomic Signatures in the O-Antigen Biosynthesis Gene Cluster.

Vibrio anguillarum is the most frequent pathogen affecting fish worldwide. The only known virulent strains of V. anguillarum are serotypes O1, O2, and O3. Genetic differences between the serotypes that could shed insight on the evolution and serotype differences of this marine pathogen are unknown. Here, we fully sequenced and characterized a strain of V. anguillarum O1 (J382) isolated from winter steelhead trout (Oncorhynchus mykiss irideus) in British Columbia, Canada. Koch's postulates using the O1 strain were replicated in naïve lumpfish (Cyclopterus lumpus) and compared to O2. Phenotypic and genotypic comparisons were conducted for serotypes O1, O2, and O3, using biochemical tests and bioinformatic tools, respectively. The genome of V. anguillarum O1 (J382) contains two chromosomes (3.13 Mb and 1.03 Mb) and two typical pJM1-like plasmids (65,573 and 76,959 bp). Furthermore, V. anguillarum O1 (J382) displayed resistance to colistin sulphate, which differs from serotype O2 and could be attributed to the presence of the ugd gene. Comparative genomic analysis, among the serotypes, showed that intra-species evolution is driven by insertion sequences, bacteriophages, and a different repertoire of putative ncRNAs. Genetic heterogeneity in the O-antigen biosynthesis gene cluster is characterized by the absence or the presence of unique genes, which could result in differences in the immune evasion mechanisms employed by the respective serotypes. This study contributes to understanding the genetic differences among V. anguillarum serovars and their evolution.

Accidentes vinculados a la Cirugía Plástica Pediátrica durante el confinamiento voluntario en Uruguay por SARS-CoV-2 / Accidents related to Pediatric Plastic Surgery during voluntary confinement in Uruguay due to SARS-CoV-2

Introducción y objetivo: Epidemiológicamente se esperaban cambios en los patrones de consulta en emergencia durante la pandemia por SARS-CoV-2. La permanencia dentro del hogar de todos los integrantes de la familia, la suspensión de la actividad deportiva, recreativa y social, sumadas al temor por concurrir a emergencia y contraer COVID-19 podían generar la interrogante en cuanto a la incidencia de accidentes. Material y método: Analizamos las historias clínicas de las consultas en emergencia de 3 instituciones en Uruguay, país en el cual el confinamiento durante la pandemia por COVID-19 fue voluntario desde el 13 de marzo de 2020 al 29 de junio del mismo año. Incluimos todos los motivos de consulta vinculados a Cirugía Plástica Pediátrica en nuestro medio en pacientes menores de 16 años. Resultados: Obtuvimos datos de 166 pacientes. Durante el mismo período del 2019 se recabaron datos de 268 pacientes. Conclusiones: Nuestro trabajo evidencia menos accidentes vinculados a Cirugía Plástica Pediátrica durante el confinamiento. Los pacientes de sexo masculino fueron los más afectados durante el confinamiento y fuera de él. Hubo una disminución de accidentes en pacientes de 6 a 16 años, siendo la franja más afectada la de los pacientes de edad prescolar. Aumentaron las mordeduras de perro, disminuyeron las consultas menos graves junto con los procedimientos más sencillos y no se evidenció aumento en los ingresos hospitalarios ni en las cirugías por accidentes. (AU)

Background and objective: Changes in the epidemiology at the emergency department during the SARS-CoV-2 pandemic were expected. Confinement of all family members, the suspension of sports, and social activity, in addition to the fear of attending an emergency and contracting COVID-19 could change the incidence of accidents. Methods: The medical records of the emergency department of 3 institutions in Uruguay, country in which confinement during the COVID-19 pandemic was voluntary from March 13, 2020 to June 29 of the same year. All the consults related to Pediatric Plastic Surgery in patients under 16 years of age were included. Results: Data was obtained from 166 patients during the period of voluntary confinement. During the same period of 2019, data from 268 patients were collected. Conclusions: In our study, fewer accidents related to Pediatric Plastic Surgery were present during confinement. Male patients were the most affected in both periods. A decrease in accidents was evidenced in patients aged 6-16 years, with preschool-age patients being the most affected group. Not severe consultations decreased along with the simplest procedures and there were no increases in hospital admissions or surgeries due to accidents. (AU)

Relative expression and validation of Aeromonas salmonicida subsp. salmonicida reference genes during ex vivo and in vivo fish infection.

The genus Aeromonas is found worldwide in freshwater and marine environments and has been implicated in the etiology of human and animal diseases. In fish, among Aeromonas species, A. salmonicida causes massive mortality and great economic losses in marine and continental aquaculture species. Currently, several aspects of the clinical signs and pathogenesis of this Gram-negative bacterium have been described; however, determination of an appropriate reference gene is essential to normalize cellular mRNA data remain unknown. Here we evaluate the stability of seven candidate reference genes to be used for data normalization during ex vivo and in vivo experiments conducted in Atlantic cod, Atlantic salmon, and lumpfish. To assess this, raw Ct values obtained were evaluated by using geNorm, NormFinder, BestKeeper, Delta Ct comparison, and the comprehensive ranking, through the bioinformatic open-access portal RefFinder. We determined that fabD and era were most suitable reference genes in Atlantic cod primary macrophages, hfq and era in Atlantic salmon primary macrophages, rpoB and fabD in lumpfish head kidney samples, and hfq and era in lumpfish spleen. Our study demonstrates that use of multiple reference genes and its validation before measurements helps to minimize variability arising in qPCR studies that evaluate A. salmonicida gene expression in fish tissues. Overall, this study provided with an expanded list of reliable reference genes for A. salmonicida gene expression using qPCR during fish infection studies.

Simulación in situ y "madre" simulada, nuevas estrategias pedagógicas en pediatría / Simulação in situ e simulação "mãe", novas estratégias pedagógicas em pediatria / In situ simulation and simulated "mother", new pedagogical strategies in pediatrics

El proceso de docencia-aprendizaje de las disciplinas clínicas tiene entre sus desafíos la adquisición de habilidades del estudiante con el paciente. Las oportunidades de prácticas clínicas se han reducido. La menor cantidad de consultas pediátricas en el año 2020 durante la pandemia por Coronoavirus exacerbó este hecho. Objetivo: Comunicar la experiencia de simulación in situ y madre simulada realizada por docentes del Departamento de Emergencia Pediátrica del Centro Hospitalario Pereira Rossell, dirigida al entrenamiento de estudiantes de la Unidad Curricular Pediatría (Facultad de Medicina-UdelaR), en la realización de la anamnesis de un paciente con patología respiratoria. Metodología: estudio descriptivo. Madre simulada: docentes/residentes. Simulador: lactante-tecnología intermedia. Participantes del escenario: 2 estudiantes. Lugar: DEP-CHPR. Período: setiembre-diciembre 2020. Resultados: Participaron de la simulación 327 estudiantes, 255 contestaron el censo. Utilidad de la simulación: 53,2% muy útil/excelente, poco o nada útil 24,8%, no contesta 22%. Credibilidad: aceptable 38%, muy creíble/excelente 27,5%, 12,5% poco o nada creíble, 22% no contesta. Participación grupal en el debriefing 64,5%, participación parcial 11,4%, sin debriefing 2,1%, no contesta 22%.Discusión: De los 327 estudiantes que realizaron la simulación al menos 24,% la percibió como poco o nada útil. La falta de información previa, la participación en el escenario de 2 estudiantes por grupo y la poca interacción en el debriefing en algunos grupos pudo haber influído. Conclusiones: La experiencia comunicada generó nuevas oportunidades de docencia aprendizaje. Se identificaron aspectos metodológicos que pueden mejorarse.

The teaching-learning process in clinical disciplines includes amongst its challenges the acquisition of the student's skills with the patient. Opportunities for clinical practice have diminished. A smaller number of pediatric visits in 2020 during the Coronavirus pandemic increased this fact. Objective: to communicate the experience of in situ simulation and simulated mother carried out by the faculty of the Pediatric Emergency Department of the Pereira Rossell Hospital Center, address to students in the Pediatrics Curricular Unit (School of Medicine- UdelaR), in the case of an infant patient with respiratory pathology. Methods: Descriptive study. Simulated mother: faculty members/residents. Simulator: intermediate technology infant. Participants in the scenario: 2 students. Location: DPE.PRHC. Period: September- December 2020.Results: 327 students participated in the simulation, 255 answered the census. Usefulness of the simulation: 53.2% very useful / excellent, little or not at all useful 24.8%, no answer 22%. Credibility: acceptable 38%, very credible / excellent 27.5%, 12.5% ​​little or not at all credible, 22% do not answer. Group participation in the debriefing 64.5%, partial participation 11.4%, without debriefing 2.1%, no answer 22%. Discussion: Of the 327 students who carried out the simulation at least 24,8 % perceived it as little or not useful. The lack of previous information, the participation in the scenario of 2 students per group and the little interaction in the debriefing in some groups could have influenced. Conclusions: The communicated experience generated new teaching-learning opportunities. Methodological aspects that can be improved were identified.

O processo de ensino-aprendizagem de disciplinas clínicas tem entre seus desafios a aquisição de habilidades do aluno com o paciente. As oportunidades de estágio clínico foram reduzidas. O menor número de consultas pediátricas em 2020 durante a pandemia de Coronoavirus exacerbou esse fato. Objetivo: Comunicar a experiência de simulação in situ e simulação materna realizada pela docentes do Serviço de Urgência Pediátrica do Centro Hospitalar Pereira Rossell, com alunos da Unidade Curricular Pediatria (Faculdade de Medicina-UdelaR), na anamnese de paciente com doença respiratória. Metodologia: estudo descritivo. Mãe simulada: professores / residentes. Simulador: lactente de tecnologia intermédia. Participantes do cenário: 2 alunos. Local: DEP-CHPR. Período: setembro a dezembro de 2020. Resultados: 327 alunos participaram da simulação, 255 responderam a pesquisa. Utilidade da simulação: 53,2% muito útil / excelente, pouco ou nada útil 24,8%, sem resposta 22%. Credibilidade: aceitável 38%, muito credível / excelente 27,5%, 12,5% pouco ou nada credível, 22% não respondem. Participação do grupo no debriefing 64,5%, participação parcial 11,4%, sem debriefing 2,1%, sem resposta 22%. Discussão: Dos 327 alunos que realizaram a simulação, pelo menos 24,% a perceberam como pouco ou nada útil. A falta de informação prévia, a participação no cenário de 2 alunos por grupo e a pouca interação no debriefing em alguns grupos podem ter influenciado. Conclusões: A experiência comunicada gerou novas oportunidades de ensino-aprendizagem. Aspectos metodológicos que podem ser melhorados foram identificados.

Estimating Occupancy Levels in Enclosed Spaces Using Environmental Variables: A Fitness Gym and Living Room as Evaluation Scenarios.

The understanding of occupancy patterns has been identified as a key contributor to achieve improvements in energy efficiency in buildings since occupancy information can benefit different systems, such as HVAC (Heating, Ventilation, and Air Conditioners), lighting, security, and emergency. This has meant that in the past decade, researchers have focused on improving the precision of occupancy estimation in enclosed spaces. Although several works have been done, one of the less addressed issues, regarding occupancy research, has been the availability of data for contrasting experimental results. Therefore, the main contributions of this work are: (1) the generation of two robust datasets gathered in enclosed spaces (a fitness gym and a living room) labeled with occupancy levels, and (2) the evaluation of three Machine Learning algorithms using different temporal resolutions. The results show that the prediction of 3-4 occupancy levels using the temperature, humidity, and pressure values provides an accuracy of at least 97%.

Intermediary metabolic response and gene transcription modulation on the Sub-Antarctic notothenioid Eleginops maclovinus (Valenciennes, 1930) injected with two strains of Piscirickettsia salmonis.

Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic-resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α-amino acid levels increased in fish injected with AUSTRAL-005 strain compared to sham-injected and LF-89-inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α-amino acid metabolism in muscle after injection with AUSTRAL-005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic-resistant strain AUSTRAL-005.

[Left Ventricular Mixoma: A case report]. / Mixoma Ventricular Izquierdo.

Myxoma is a primary cardiac tumor of low prevalence in the population, with a predilection for the left atrium followed by the right atrium. The location in the ventricles is infrequent and the symptoms depends of the place of origin. Its associated complications and its particular form of debut can be avoided with timely surgical resection.We present the case of a 15-year-old symptomatic woman with a mass in the left ventricle. The mass was surgically resected, and the histopathological study confirmed a myxoma.

Intoxicación aguda no intencional grave con alcohol isopropílico en el Departamento de Emergencia Pediátrica del Centro Hospitalario Pereira Rossell. A propósito de tres casos clínicos / Acute unintentional poisoning with isopropyl alcohol in the Pereira Rossell Pediatric Emergency Ward. Three clinical cases / Intoxicação aguda não intencional com álcool isopropílico na Emergência Pediátrica do Centro Hospitalar Pereira Rossell. Três relatos de caso

Resumen: Las intoxicaciones agudas en los servicios de emergencia pediátrica ocupan un lugar importante dentro de la patología pediátrica de urgencia por su potencial riesgo vital. El Centro de Información y Asesoramiento Toxicológico (CIAT) recibe anualmente un promedio de 10.000 consultas, el 45% corresponde a la edad pediátrica. La intoxicación aguda por isopropanol es poco frecuente, pero potencialmente grave. El CIAT ha registrado 34 casos en los últimos cinco años. El alcohol isopropílico se encuentra en muchos productos de limpieza y desinfectantes. La intoxicación aguda se caracteriza inicialmente por síntomas digestivos. En la evolución pueden agregarse manifestaciones neurológicas: depresión de conciencia, hipotonía e hiporreflexia, asociada a hipoglicemia, gap osmolar elevado, cetonemia y cetonuria. En casos graves puede ocasionar hipotermia, hipotensión y arritmias. La acidosis metabólica es poco frecuente, a diferencia de otros alcoholes. En este caso, la toxicidad aguda es dependiente en gran medida de la formación de metabolitos. En este trabajo son isopropanol y acetona los responsables de las manifestaciones clínicas y alteraciones en los estudios de laboratorio. Se analizaron tres casos clínicos en niños de 96, 18 y 15 meses. Los tres casos evolucionaron de forma favorable en 72-96 horas, y, como se refiere en la literatura, el pronóstico en general es bueno y sin secuelas. El abordaje de estos pacientes en la urgencia se basa en la aplicación del triángulo de evaluación pediátrica y la secuencia A, B, C, D, E con el propósito de identificar en forma precoz la necesidad de realizar correcciones metabólicas o soporte de funciones.

Summary: Acute poisoning in pediatric emergency services is an important part of the emergency pediatric pathology due to its potential risk. CIAT receives an average of 10,000 consultations annually, 45% of which are from children. Acute isopropanol poisoning is rare but potentially serious. CIAT has recorded 34 cases in the last 5 years. Isopropyl alcohol is found in many cleaning products and disinfectants. Acute poisoning is initially characterized by digestive symptoms. Some neurological manifestations may arise during the acute poisoning evolution, such as, depression of consciousness, hypotonia and hyporeflexia, associated with hypoglycemia, elevated osmolar gap, ketonemia and ketoneuria. In severe cases, it can cause hypothermia, hypotension and arrhythmia. Metabolic acidosis is rare, unlike what happens with other alcohols. In this case, acute toxicity largely depends on the formation of metabolites; in this case, isopropanol and acetone are responsible for clinical manifestations and alterations in laboratory studies. We hereby present three clinical cases of children aged 96, 18 and 15 months. The three cases evolved favorably in 72-96 hours and, as described in the literature, the prognosis in general is good and without sequelae. The approach to these patients in the emergency ward was the pediatric evaluation triangle and the A, B, C, D, E sequence in order to identify early the need to perform metabolic corrections and / or support functions.

Resumo: As intoxicações agudas nos serviços de emergência pediátrica ocupam um lugar importante na patologia pediátrica de emergência devido ao seu risco potencial para a vida. O CIAT recebe, em média, 10.000 consultas anualmente, 45% de pacientes pediátricos. A intoxicação aguda por isopropanol é rara, mas potencialmente grave. O CIAT registrou 34 casos nos últimos 5 anos. O álcool isopropílico encontra-se em vários produtos de limpeza e desinfetantes. O envenenamento agudo é inicialmente caracterizado por sintomas digestivos. Na evolução do envenamento, podem aparecer manifestações neurológicas: depressão da consciência, hipotonia e hiporreflexia, associadas à hipoglicemia, gap osmolar elevado, cetonemia e cetoneúria. Em casos graves, pode causar hipotermia, hipotensão e arritmias. A acidose metabólica é rara, diferentemente do que acontece com outros álcoois. Nesse caso, a toxicidade aguda depende em grande parte da formação de metabólitos; neste caso, isopropanol e acetona são responsáveis pelas manifestações clínicas e alterações em estudos laboratoriais. Analisaram-se três casos clínicos em crianças de 96, 18 e 15 meses. Os três casos evoluíram favoravelmente em 72-96 horas e, como relatamos na literatura, o prognóstico em geral é bom e sem sequelas. A abordagem desses pacientes na emergência foi a aplicação do triângulo de avaliação pediátrica na sequência A, B, C, D, E, a fim de identificar precocemente a necessidade de realizar correções metabólicas e / ou suporte de funções.

Aeromonas salmonicida infects Atlantic salmon (Salmo salar) erythrocytes.

Aeromonas salmonicida subsp. salmonicida (hereafter A. salmonicida) is the aetiological agent of furunculosis in marine and freshwater fish. Once A. salmonicida invade the fish host through skin, gut or gills, it spreads and colonizes the head kidney, liver, spleen and brain. A. salmonicida infects leucocytes and exhibits an extracellular phase in the blood of the host; however, it is unknown whether A. salmonicida have an intraerythrocytic phase. Here, we evaluate whether A. salmonicida infects Atlantic salmon (Salmo salar) erythrocytes in vitro and in vivo. A. salmonicida did not kill primary S. salar erythrocytes, even in the presence of high bacterial loads, but A. salmonicida invaded the S. salar erythrocytes in the absence of evident haemolysis. Naïve Atlantic salmon smolts intraperitoneally infected with A. salmonicida showed bacteraemia 5 days post-infection and the presence of intraerythrocytic A. salmonicida. Our results reveal a novel intraerythrocytic phase during A. salmonicida infection.

Aeromonas salmonicida subsp. salmonicida Early Infection and Immune Response of Atlantic Cod (Gadus morhua L.) Primary Macrophages.

In contrast to other teleosts, Atlantic cod (Gadus morhua) has an expanded repertoire of MHC-I and TLR components, but lacks the MHC-II, the invariant chain/CD74, and CD4+ T cell response, essential for production of antibodies and prevention of bacterial infectious diseases. The mechanisms by which G. morhua fight bacterial infections are not well understood. Aeromonas salmonicida subsp. salmonicida is a recurrent pathogen in cultured and wild fish, and has been reported in Atlantic cod. Macrophages are some of the first responders to bacterial infection and the link between innate and adaptive immune response. Here, we evaluated the viability, reactive oxygen species (ROS) production, cell morphology, and gene expression of cod primary macrophages in response to A. salmonicida infection. We found that A. salmonicida infects cod primary macrophages without killing the cod cells. Likewise, infected Atlantic cod macrophages up-regulated key genes involved in the inflammatory response (e.g., IL-1ß and IL-8) and bacterial recognition (e.g., BPI/LBP). Nevertheless, our results showed a down-regulation of genes related to antimicrobial peptide and ROS production, suggesting that A. salmonicida utilizes its virulence mechanisms to control and prevent macrophage anti-bacterial activity. Our results also indicate that Atlantic cod has a basal ROS production in non-infected cells, and this was not increased after contact with A. salmonicida. Transmission electron microscopy results showed that A. salmonicida was able to infect the macrophages in a high number, and release outer membrane vesicles (OMV) during intracellular infection. These results suggest that Atlantic cod macrophage innate immunity is able to detect A. salmonicida and trigger an anti-inflammatory response, however A. salmonicida controls the cell immune response to prevent bacterial clearance, during early infection.

Identification and validation of reliable Aeromonas salmonicida subspecies salmonicida reference genes for differential gene expression analyses.

Aeromonas salmonicida subsp. salmonicida is a Gram-negative, facultative intracellular pathogen of a wide range of freshwater and marine fish species. A. salmonicida is the causative agent of furunculosis, an immunosuppressive disease that typically progresses to septicemia. Several aspects of A. salmonicida pathogenesis has already been described, but fundamental genetic aspects of the psychrophilic lifestyle of this bacterium remain unknown. Reverse transcription quantitative real-time polymerase chain reaction (qPCR) is a precise molecular technique used to detect very slight changes in gene expression. The appropriate choice of reference genes is essential for accurate normalization of qPCR gene expression data. Despite the available abundance of validated reference genes for mesophilic pathogens, a broad list of validated reference genes for A. salmonicida is not available. Here, we evaluated seven A. salmonicida reference genes under different culture conditions, including different growth phases, iron-limited and iron-supplemented conditions, and thermal stress. We determined that hfq maintained the most stable expression, followed by era, recA, rpoB, 16S, fabD, and gapA. The results of this study provided with an expanded list of reliable reference genes for A. salmonicida gene expression studies using qPCR.

Risk factors and predisposing conditions for urinary tract infection.

Understanding individual and population-specific risk factors associated with recurrent urinary tract infections (UTIs) can help physicians tailor prophylactic strategies. Frequent intercourse, vulvovaginal atrophy, change of the local bacterial flora, history of UTIs during premenopause or in childhood, family history, and a nonsecretor blood type are substantiated risk factors for recurrent uncomplicated UTIs. This is a narrative review based on relevant literature according to the experience and expertise of the authors. Asymptomatic bacteriuria is generally benign; however, during pregnancy it is more common and is associated with an increased likelihood of symptomatic infection, which may harm the mother or fetus. Screening of pregnant women and appropriate treatment with antimicrobials must be balanced with the potential for adverse treatment-related outcomes; appropriate prophylaxis should be considered where possible. High-quality data are currently lacking on risks related to asymptomatic bacteriuria in pregnancy and further data in this hard-to-study population should be a primary concern for researchers. Incomplete voiding represents the primary risk factor for UTIs associated with conditions such as urinary incontinence and prolapse. Correcting the presence of residual urine remains the most effective prophylaxis in these populations. Bladder function alters throughout life; however, changes in function may be particularly profound in clinical populations at high risk of UTIs. Patients with neurogenic bladder will also likely have other evolving medical issues which increase the risk of UTIs, such as repeated catheterization and increasing residual urine volume. More aggressive antimicrobial prophylactic strategies may be appropriate in these patients. Again, the paucity of data on prophylaxis in these high-risk patients requires the attention of the research community.

Effects of Vitamin D2 (Ergocalciferol) and D3 (Cholecalciferol) on Atlantic Salmon (Salmo salar) Primary Macrophage Immune Response to Aeromonas salmonicida subsp. salmonicida Infection.

Vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol) are fat-soluble secosteroid hormones obtained from plant and animal sources, respectively. Fish incorporates vitamin D2 and D3 through the diet. In mammals, vitamin D forms are involved in mineral metabolism, cell growth, tissue differentiation, and antibacterial immune response. Vitamin D is an essential nutrient in aquafeeds for finfish. However, the influence of vitamin D on fish cell immunity has not yet been explored. Here, we examined the effects of vitamin D2 and vitamin D3 on Salmo salar primary macrophage immune response to A. salmonicida subspecies salmonicida infection under in vitro conditions. We determined that high concentrations of vitamin D2 (100,000 ng/ml) and D3 (10,000 ng/ml) affect the growth of A. salmonicida and decrease the viability of S. salar primary macrophages. In addition, we determined that primary macrophages pre-treated with a biologically relevant concentration of vitamin D3 for 24 h showed a decrease of A. salmonicida infection. In contrast, vitamin D2 did not influence the antibacterial activity of the S. salar macrophages infected with A. salmonicida. Vitamin D2 and D3 did not influence the expression of canonical genes related to innate immune response. On the other hand, we found that A. salmonicida up-regulated the expression of several canonical genes and suppressed the expression of leukocyte-derived chemotaxin 2 (lect-2) gene, involved in neutrophil recruitment. Primary macrophages pre-treated for 24 h with vitamin D3 counteracted this immune suppression and up-regulated the transcription of lect-2. Our results suggest that vitamin D3 affects A. salmonicida attachment to the S. salar primary macrophages, and as a consequence, the A. salmonicida invasion decreased. Moreover, our study shows that the positive effects of vitamin D3 on fish cell immunity seem to be related to the lect-2 innate immunity mechanisms. We did not identify positive effects of vitamin D2 on fish cell immunity. In conclusion, we determined that the inactive form of vitamin D3, cholecalciferol, induced anti-bacterial innate immunity pathways in Atlantic salmon primary macrophages, suggesting that its utilization as a component of a healthy aquafeed diet in Atlantic salmon could enhance the immune response against A. salmonicida.

Draft Genome Sequence of the Type Strain Aeromonas salmonicida subsp. salmonicida ATCC 33658.

Here, we report the draft genome sequence of the type strain Aeromonas salmonicida subsp. salmonicida ATCC 33658 isolated from Salmo salar The size of the genome is 4,728,143 bp with a G+C content of 58.5%. The A. salmonicida subsp. salmonicida ATCC 33658 genome lacks essential virulence genes that were likely lost during genomic rearrangements.

Epidemiology and outcomes of dermatology in-patient consultations in a Midwestern U.S. university hospital.

CONTEXT: A paucity of data exists concerning the utilization of in-patient dermatologic consultations. Previous studies on this subject have indicated a knowledge deficit of primary care providers with regard to common dermatoses, prompting a need for more effective teaching mechanisms in this area. OBJECTIVE: To identify dermatologic conditions in the in-patient setting that are frequently misdiagnosed by non-dermatologists in order to improve future patient care and cost reduction through physician education. DESIGN: Retrospective chart review of 271 consecutive dermatologic consultations from primary ward teams between January 20, 1998, and May 19, 1999. SETTING: Non-dermatology in-patient services at a Midwestern state-supported university hospital system in the U.S. PATIENTS: Patients hospitalized on non-dermatology wards with skin problems prompting a formal dermatologic consultation. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Prevalence of dermatologic conditions that are most frequently misdiagnosed on non-dermatology in-patient services. RESULTS: Seventy-six percent of the dermatologic consults were requested by Internal Medicine, Surgery, and Psychiatry departments. Common skin conditions accounted for a large majority of dermatologic consultations including: dermatitis (21.0%) and drug eruption (10.0%). The primary ward team submitted a correct dermatologic diagnosis in only 23.9 percent of cases. Dermatology consultation resulted in a change in or addition to treatment in 77 percent of patients. CONCLUSIONS: Our results suggest that common skin conditions account for a large majority of dermatologic consultations in a University hospital setting. Modern hospital ward teams continue to struggle with accurately recognizing and appropriately managing common skin problems resulting in inappropriate treatment, wasted resources, and prolonged hospitalization. Increasing medical student and house staff knowledge and experience in the diagnosis and management of common skin disorders could help address this problem.