Cost-effective fully 3D-printed on-drop electrochemical sensor based on carbon black/polylactic acid: a comparative study with screen-printed sensors in food analysis.

3D-printing technology allows scientist to fabricate easily electrochemical sensors. Until now, these sensors were designed employing a large amount of material, which increases the cost and decreases manufacturing throughput. In this work, a low-cost 3D-printed on-drop electrochemical sensor (3D-PES) was fully manufactured by fused filament fabrication, minimizing the number of printing layers. Carbon black/polylactic acid filament was employed, and the design and several printing parameters were optimized to yield the maximum electroanalytical performance using the minimal amount of material. Print speed and extrusion width showed a critical influence on the electroanalytical performance of 3D-PES. Under optimized conditions, the fabrication procedure offered excellent reproducibility (RSD 1.3% in working electrode diameter), speed (< 3 min/unit), and costs (< 0.01 $ in material cost). The 3D-PES was successfully applied to the determination of phloridzin in apple juice. The analytical performance of 3D-PES was compared with an equivalent commercial on-drop screen-printed electrode, yielding similar precision and accuracy but lower sensitivity. However, 3D-PES provides interesting features such as recyclability, biodegradability, low-cost, and the possibility of being manufactured near the point of need, some of which meets several demands of Green Chemistry. This cost-effective printing approach is a green and promising alternative for manufacturing disposable and portable electroanalytical devices, opening new possibilities not only in on-site food analysis but also in point-of-care testing.

Colombian consensus for the molecular diagnosis of endometrial cancer / Consenso colombiano para el diagnóstico molecular del cáncer de endometrio.

Objective: The Cancer Genome Atlas research program (TCGA) developed the molecular classification for endometrial cancer with prognostic and therapeutic utility, which was replaced by the ProMisE (Proactive Molecular Risk Classifier for Endometrial Cancer) classification by consensus and international guidelines due to its high cost. This article aims to present national recommendations from an expert consensus that allows unification and implementation of the molecular classification for women with endometrial cancer nationwide, with a rational use of resources and technology. Methods: Consensus of 36 experts in clinical oncology, oncological gynecology, pathology, and genetics, with clinical practice in the national territory. The leader group performed a literature review and structuring of questions rated 1 to 9 points. A modified nominal group technique was used. There was a face-to-face meeting with master presentations, deliberative dialogue, and Google Forms (Google LLC, Mountain View, CA, USA) questionnaire voting with analysis and discussion of responses. The non-consensual responses led to a second round of voting. The final manuscript was finally prepared and revised. Results: Seven recommendations were formulated integrating the panelist responses based on evidence, but adjusted to the Colombian context and reality. Recommendation 1. The molecular classification is recommended in all the endometrial cancers using the immunohistochemistry markers as subrogated results from the molecular profile initially proposed in the TCGA classification. Recommendation 2. The sequential test strategy is recommended, starting with the immunohistochemistry markers (p53, MLH1, MSH 2, MSH6, PMS2) simultaneously in all the patients, defining to request POLE (DNA polymerase epsilon) (if available) according to the risk classification based on the surgical piece. Recommendation 3. It is recommended, that the gynecologist oncologist should be the one to request the POLE (if available) according to the final pathology report. This test must be requested for all endometrial cancers stage I-II, except in low risk (stage IA low grade endometrioid histology without linfovascular invasion normal p53) and, stages III-IV without residual disease, without affecting the request of subrogated immunohistochemistry molecular markers upon histology. The consensus proposes that the POLE is requested after the immunohistochemistry and according to the categories in the risk classification established by the 2020 ESGO/ESTRO/ESP guidelines. Recommendation 4. It is recommended to perform immunohistochemistry for hormonal receptors for all women with endometrial cancer and the HER2 in patients with p53abn, simultaneously with the others immunohistochemistry markers. Recommendation 5. It is recommended to perform the immunohistochemistry markers (p53, MLH1, MSH2, MSH6 y PMS2) in an initial endometrial biopsy or curettage when the specimen is adequate and available. In case the initial immunohistochemistry is inconclusive, or there are histological discrepancies between the initial and definitive pathology, it is recommended to repeat the molecular profile in the surgical pathology. The immunohistochemistry markers must be reported in the pathology report according to the CAP (College of American Pathologists) recommendations, independently of the type of sample. Recommendation 6. It is recommended to perform MLH1 promoter methylation testing in patients who exhibit loss of expression of MLH1 in immunohistochemistry whether it is accompanied or not with loss of expression of PMS2. All the patients with deficient MMR (mismatch repair), should be sent for genetic counseling to rule out Lynch syndrome. Recommendation 7. It is recommended to consider the molecular classification in addition to the classical histopathological criteria when making adjuvant judgments, as incorporated by the classification of prognostic groups of the 2020 ESGO/ESTRO/ESP guidelines. Conclusions: It is necessary to implement the molecular classification of endometrial cancer in clinical practice in accordance to the Colombian context, due to its prognostic and probably predictive value. This will enable the characterization of the Colombian population in order to offer individualized guided treatments. This is an academic and nonregulatory document.

Objetivos: el programa Cancer Genome Atlas Research (TCGA) desarrolló la clasificación molecular para cáncer endometrial con utilidad pronóstica y terapéutica, la cual ha sido reemplazada por consensos y guías internacionales por la clasificación ProMisE (Proactive Molecular Risk Classifier for Endometrial Cancer) debido a su alto costo. El objetivo de este artículo es presentar recomendaciones a nivel nacional derivadas de un consenso de expertos que permitan unificar e implementar la clasificación molecular para mujeres con cáncer endometrial, mediante un uso racional de recursos y tecnología. Materiales y métodos: consenso de 36 expertos en oncología clínica, ginecología oncológica, patología y genética con práctica clínica en el territorio nacional. El grupo líder realizó una revisión de la literatura y estructuración de preguntas calificadas de 1 a 9 puntos. Se utilizó la técnica de grupo nominal modificada. Se efectuaron reuniones presenciales con presentaciones magistrales, diálogo deliberativo y votación de cuestionario Google Forms (Google LLC, Mountain View, CA, USA) con análisis y discusión de respuestas. Las respuestas no consensuadas se llevaron a una segunda ronda de votación. Finalmente, se elaboró y revisó el manuscrito final. Resultados: se formularon siete recomendaciones integrando las respuestas de las panelistas basadas en evidencia, pero ajustadas al contexto y a la realidad colombiana. Recomendación 1. Se recomienda realizar la clasificación molecular en todos los carcinomas endometriales utilizando los marcadores de inmunohistoquímica como resultados subrogados del perfil molecular inicialmente propuesto en la clasificación del TCGA. Recomendación 2. Se recomienda la estrategia secuencial de testeo iniciando por los marcadores de inmunohistoquímica (p53, MLH1, MSH 2, MSH6, PMS2) simultáneamente en todas las pacientes, y definir la solicitud del POLE (polimerasa épsilon del DNA) (si se encuentra disponible) de forma diferida de acuerdo con la clasificación de riesgo basado en la pieza quirúrgica. Recomendación 3. Se recomienda que sea el ginecólogo oncólogo quien solicite el POLE (si se encuentra disponible) de acuerdo con el reporte de patología definitivo. Esta prueba se debe solicitar a todos los cánceres endometriales de estadio I-II, excepto los de bajo riesgo (estadio IA endometrioide de bajo grado sin invasión linfovascular p53 normal) y estadio III-IV sin enfermedad residual, sin afectar la solicitud de los marcadores moleculares subrogados por inmunohistoquímica de acuerdo con la histología. El consenso propone que la solicitud del POLE se realice posterior a la inmunohistoquímica y de acuerdo con la clasificación del riesgo según las categorías establecidas por la guía ESGO/ESTRO/ESP del 2020. Recomendación 4. Se recomienda realizar simultáneamente con los otros marcadores de inmunohistoquímica la prueba para receptores hormonales en todas las pacientes con cáncer endometrial y el HER2 en pacientes con p53abn. Recomendación 5. Se recomienda que los marcadores de inmunohistoquímica (p53, MLH1, MSH2, MSH6 y PMS2) se realicen en la biopsia/legrado endometrial inicial cuando la muestra es adecuada y está disponible. En caso de inmunohistoquímica inicial no concluyente, o discrepancias histológicas entre la patología inicial y definitiva, se recomienda repetir el perfil molecular en la patología quirúrgica. Los marcadores de inmunohistoquímica deben reportarse en el informe de patología de acuerdo con las recomendaciones del CAP (College of American Pathologists), independientemente del tipo de muestra. Recomendación 6. Se recomienda realizar estudio de metilación de promotor de MLH1 en pacientes con pérdida de expresión de MLH1 en la inmunohistoquímica, acompañado o no de pérdida de expresión de PMS2. Todas las pacientes con déficit de MMR (mismatch repair), deben ser enviadas a genética para descartar síndrome de Lynch. Recomendación 7. Se recomienda tener en cuenta la clasificación molecular, además de los criterios histopatológicos clásicos para la toma de decisiones de adyuvancia, tal como los incorpora la clasificación de los grupos pronósticos de la guía ESGO/ ESTRO/ESP del 2020. Conclusiones: es necesario implementar la clasificación molecular de cáncer de endometrio en la práctica clínica acorde al contexto colombiano, dado su valor pronóstico y posiblemente predictivo. Esto permitirá la caracterización de la población colombiana para ofrecer tratamientos guiados de manera individualizada. Se trata de un documento académico y no regulatorio.

Presentation and Management of Uterine Didelphys with Unilateral Cervicovaginal Agenesis/Dysgenesis (CVAD): A Multicenter Case Series.

INTRODUCTION: There are several well-described presentations of uterine didelphys (UD): UD without vaginal septum, UD with non-obstructed longitudinal vaginal septum, or UD with duplicated vaginas and an obstructed hemivagina on one side with ipsilateral renal anomaly. STUDY OBJECTIVE: To describe another variant of UD and compare the presentation and management across different institutions METHODS: This was a retrospective case series approved by the NASPAG Fellows Research Consortium. Participating institutions obtained IRB approval. Inclusion criteria included a diagnosis of UD and unilateral cervicovaginal agenesis/dysgenesis (CVAD). Descriptive statistics were used. RESULTS: Five patients met the inclusion criteria, with ages ranging from 13 to 27 years. Presenting symptoms included dysmenorrhea (80%), irregular bleeding (40%), acute onset left lower quadrant pain (20%), and abdominal mass (20%). Three patients had additional known abnormalities, including solitary kidney and solitary adrenal gland. All patients underwent pelvic magnetic resonance imaging. Two cases were only suspicious for unilateral CVAD on imaging and required pathology review postoperatively to confirm diagnosis. Two cases required a 2-staged approach with an initial diagnostic surgery followed by a second definitive procedure. Three patients were noted to have endometriosis intraoperatively. Postoperative follow-up ranged from 2 months to 2 years, with 1 patient reporting chronic pelvic pain. CONCLUSION: Diagnosis on the basis of pelvic imaging can be difficult, as this unique variant may mimic classic obstructed hemivagina with ipsilateral renal anomaly. In patients with UD with unilateral CVAD, standard management is removal of the obstructed uterine horn. This multicenter series stresses awareness about the clinical presentation, distinguishes cases of cervical agenesis from dysgenesis, and reviews approaches to management.

Genetic diagnosis of Duchenne and Becker muscular dystrophy through mRNA analysis: new splicing events.

BACKGROUND: Up to 7% of patients with Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD) remain genetically undiagnosed after routine genetic testing. These patients are thought to carry deep intronic variants, structural variants or splicing alterations not detected through multiplex ligation-dependent probe amplification or exome sequencing. METHODS: RNA was extracted from seven muscle biopsy samples of patients with genetically undiagnosed DMD/BMD after routine genetic diagnosis. RT-PCR of the DMD gene was performed to detect the presence of alternative transcripts. Droplet digital PCR and whole-genome sequencing were also performed in some patients. RESULTS: We identified an alteration in the mRNA level in all the patients. We detected three pseudoexons in DMD caused by deep intronic variants, two of them not previously reported. We also identified a chromosomal rearrangement between Xp21.2 and 8p22. Furthermore, we detected three exon skipping events with unclear pathogenicity. CONCLUSION: These findings indicate that mRNA analysis of the DMD gene is a valuable tool to reach a precise genetic diagnosis in patients with a clinical and anatomopathological suspicion of dystrophinopathy that remain genetically undiagnosed after routine genetic testing.

Adecuación al flujograma PROA de neumonía aguda comunitaria en la emergencia del Hospital de Clínicas en 2019, Uruguay / Adaptation to the PROA flowchart for community acute pneumonia in the Hospital de Clínicas emergency in 2019, Uruguay / Adaptação ao fluxograma PROA para pneumonia aguda comunitária na emergência do Hospital de Clínicas em 2019, Uruguai

Resumen: Introducción: La resistencia antimicrobiana es uno de los principales problemas de salud pública mundial. Representa una causa importante de morbilidad en la población general y un elevado costo para los sistemas sanitarios. La Neumonía Aguda Comunitaria (NAC) representa una de las principales infecciones bacterianas en nuestro medio. Objetivo general: Evaluar la adecuación al flujograma del Programa de Optimización de Antimicrobianos (PROA) para el manejo de NAC en Departamento de Emergencia del Hospital de Clínicas (HC) entre julio y agosto de 2019. Materiales y métodos: Se realizó un estudio observacional, transversal, en el período de julio-agosto de 2019, en Departamento de Emergencia del Hospital de Clínicas. Se incluyeron pacientes mayores de 18 años, que firmaron el consentimiento informado, diagnosticados con NAC, cumpliendo criterios clínicos e imagenológicos establecidos en el flujograma del PROA del Hospital de Clínicas. Se elaboró una base de datos diseñada a partir del flujograma. Resultados: Se incluyeron 51 pacientes para el análisis. La edad promedio fue 54 años, 28 eran mujeres. Las comorbilidades más prevalentes fueron: tabaquismo, consumo de pasta base de cocaína y alcoholismo, presentes en 51% de la muestra. Treinta y cinco pacientes presentaron criterios de severidad, predominando insuficiencia respiratoria en 71%. Un 43 % presentaron factores de riesgo para microorganismos multirresistentes. Se observó una adecuación al PROA de 41%. Discusión: La adecuación al tratamiento recomendado fue inferior a la descrita en otros trabajos. El principal problema fue una errónea clasificación en los grupos de riesgo propuestos en el flujograma, ocasionando la hospitalización de pacientes que debieron recibir tratamiento ambulatorio, recibiendo antibioticoterapia de mayor espectro. Conclusiones: La existencia de PROA hospitalarios permite realizar monitoreo de prácticas diagnósticas y prescripción de antimicrobianos. Se observó una inadecuada aplicación del flujograma, lo que determinó el uso de antibióticos de mayor espectro con riesgo potencial del desarrollo de resistencia.

Abstract: Introduction: Antimicrobial resistance is one of the main world public health problems. It represents an important cause of morbidity in general population and a high cost for health systems. Community Acquired Pneumonia (CAP) represents one of the main bacterial infections in our midst. Objective: To evaluate the adequacy of the Antimicrobial Stewardship (AMS) in the management of CAP in the Emergency Department of Hospital de Clínicas (HC) between July and August 2019. Materials and methods: An observational, cross-sectional study was conducted from July-August 2019, in the Emergency Department of Hospital de Clínicas. Patients older than 18 years old were included, who signed the informed consent, diagnosed with CAP, fulfilling clinical and imaging criteria established in the flowchart. A database designed from the AMS flow chart of the Hospital de Clínicas was developed. Results: 51 patients were included for the analysis. The average age was 54 years, 28 were women. The most prevalent comorbidities were smoking, consumption of cocaine paste or alcoholism, present in 51% of the sample. Thirty-five patients presented severity criteria, prevailing respiratory failure in 71%. Risk factors for multiresistant microorganisms was 43%. PROA adequacy of 41.2% was observed. Discussion: The adequacy to the recommended treatment was lower than that described in other papers. The main problem was an erroneous classification in the risk groups proposed in the flowchart, causing hospitalization of patients who had to receive treatment at home, receiving broader spectrum antibiotic therapy. Conclusions: The existence of hospital stewardships allows monitoring of diagnostic practices and antimicrobial prescription. Inadequate application of the flow chart was observed, which determined the use of broader spectrum antibiotics with potential risk of developing resistance.

Resumo: Introdução: A resistência antimicrobiana é um dos principais problemas de saúde pública global. Representa uma das principais causas de morbidade na população em geral e um alto custo para os sistemas de saúde. A Pneumonia Aguda Comunitária (PAC) representa uma das principais infecções bacterianas em nosso meio. Objetivo: Avaliar a adequação do fluxograma do Programa de Otimização de Antimicrobianos (PROA) para o gerenciamento do PAC no Pronto Atendimento do Hospital de Clínicas (HC) entre julho e agosto de 2019. Materiais e métodos: Foi realizado um estudo observacional, transversal, no período de julho a agosto de 2019, no Pronto-Socorro do Hospital de Clínicas. Foram incluídos pacientes maiores de 18 anos, que assinaram o termo de consentimento livre e esclarecido, com diagnóstico de PAC, que preenchessem os critérios clínicos e de imagem estabelecidos no fluxograma do PROA do Hospital de Clínicas. Um banco de dados projetado a partir do fluxograma foi desenvolvido. Resultados: 51 pacientes foram incluídos para análise. A idade média era de 54 anos, 28 eram mulheres. As comorbidades mais prevalentes foram: tabagismo, consumo de pasta base de cocaína e etilismo, presentes em 51% da amostra. Trinta e cinco pacientes apresentaram critérios de gravidade, predominando insuficiência respiratória em 71%. 43% apresentaram fatores de risco para microrganismos multirresistentes. Observou-se adequação ao PROA de 41%. Discussão: A adequação ao tratamento recomendado foi inferior ao descrito em outros estudos. O principal problema era uma classificação errônea nos grupos de risco propostos no fluxograma, ocasionando a internação de pacientes que precisavam receber tratamento ambulatorial, recebendo antibioticoterapia de maior espectro. Conclusões: A existência de PROAs hospitalares permite o monitoramento das práticas diagnósticas e prescrição de antimicrobianos. Observou-se uma aplicação inadequada do fluxograma, que determinou o uso de antibióticos de maior espectro e com potencial risco de desenvolvimento de resistência.

Toxicity Bioassay and Cytotoxic Effects of the Benthic Marine Dinoflagellate Amphidinium operculatum.

Benthic dinoflagellates produce a wide array of bioactive compounds, primarily polyketides, that cause toxic effects on human consumers of seafood and perhaps mediate species interactions in the benthic microenvironment. This study assesses toxic and other bioactive effects of the benthic dinoflagellate Amphidinium operculatum (strain AA60) in two targeted bioassays. The brine shrimp (Artemia salina) bioassay revealed lethal effects of direct exposure to live dinoflagellate cells (Treatment A) and even higher potency with ethanolic extracts of lysed cells (Treatment D). There were no inimical bioactive effects of components released to the aqueous growth medium (Treatment B) or from aqueous cell lysates (Treatment C). The hypothesis that released bioactive compounds provide a chemical defense against metazoan grazers is therefore not supported by these results. The cytotoxic effect of ethanolic crude extracts of this dinoflagellate exhibited mild to high growth reduction effects on six human cancer cell lines. In particular, crude cell-free extracts proved highly growth-inhibitory activity towards breast and lung cancer cell lines MCF-7 and SKLU-1, respectively. Preliminary anti-cancer results indicate that natural bioactive compounds from Amphidinium are worthy of structural characterization and further toxicological investigation as potential therapeutants.

Prediction of Mechanical Properties by Artificial Neural Networks to Characterize the Plastic Behavior of Aluminum Alloys.

In metal forming, the plastic behavior of metallic alloys is directly related to their formability, and it has been traditionally characterized by simplified models of the flow curves, especially in the analysis by finite element simulation and analytical methods. Tools based on artificial neural networks have shown high potential for predicting the behavior and properties of industrial components. Aluminum alloys are among the most broadly used materials in challenging industries such as aerospace, automotive, or food packaging. In this study, a computer-aided tool is developed to predict two of the most useful mechanical properties of metallic materials to characterize the plastic behavior, yield strength and ultimate tensile strength. These prognostics are based on the alloy chemical composition, tempers, and Brinell hardness. In this study, a material database is employed to train an artificial neural network that is able to make predictions with a confidence greater than 95%. It is also shown that this methodology achieves a performance similar to that of empirical equations developed expressly for a specific material, but it provides greater generality since it can approximate the properties of any aluminum alloy. The methodology is based on the usage of artificial neural networks supported by a big data collection about the properties of thousands of commercial materials. Thus, the input data go above 2000 entries. When the relevant information has been collected and organized, an artificial neural network is defined, and after the training, the artificial intelligence is able to make predictions about the material properties with an average confidence greater than 95%.

Reliability-Based Evaluation of the Suitability of Polymers for Additive Manufacturing Intended for Extreme Operating Conditions.

A reliability engineering program must be implemented from the conceptual phase of the physical asset to define the performance requirements of the components and equipment. Thus, in this work, the aim is to find the most optimal solution to manufacture polymer-based parts for the nuclear power industry using additive manufacturing routes. This case study application has been selected because polymers processed by additive manufacturing (AM) can be well suited for nuclear applications. The methodology includes-firstly-an analysis of the suitability of materials based on high-temperature resistance, thermal aging and irradiation tolerance, considering operation conditions. Secondly, an analysis of materials' processability considering their associated AM routes is performed based on thermal analysis and evaluation of physical properties of materials. A final assessment integrating the in-service suitability and AM processability is performed using a reliability approach, solving different emerging objective conflicts through defined constraints and selection criteria. According to the integrated in-service performance evaluation: Polypropylene-ethylene polyallomer (PPP), Epoxy (EP), Phenolics (Ph), Polyurethane (PU) and Acrylonitrile butadiene rubber (NBR) are the best options for mild operation conditions and EP, Ph and PU, considering high temperature along with radiation exposure. Considering AM techniques: EP and Ph can be manufactured using VAT photopolymerization-stereolithography (VP-SLA) with a good expected processability being these materials valid for high temperature environments. Consequently, this research work analyzes the viability, processability and in-service behavior of parts.

Carey-Fineman-Ziter Syndrome: A MYMK-Related Myopathy Mimicking Brainstem Dysgenesis.

Carey-Fineman-Ziter syndrome is a congenital myopathy associated with mutations in the MYMK gene. It is clinically defined by the combination of hypotonia, Moebius-Robin sequence, facial anomalies and motor delay. Historically it was considered a brainstem dysgenesis syndrome. We provide detailed information of a Spanish boy with compound heterozygous variants in MYMK gene. A muscle biopsy performed as a toddler only disclosed minimal changes, but muscle MRI showed severe fatty infiltration of gluteus muscles and to a lesser extent in adductors magnus, sartorius and soleus muscles. Clinical course is fairly static, but the identification of new well characterized genetic cases will help to delineate the complete phenotype.

Salmonella spp. prevalence, antimicrobial resistance and risk factor determination in Colombian swine farms / Salmonella spp. prevalência, resistência antimicrobiana e determinação de fatores de risco em granjas suínas colombianas

To determine Salmonella spp. prevalence/seroprevalence, antimicrobial resistance patterns and risk factor identification associated with its presence in Colombian swine farms. 504 samples (Faeces, swabs and environment samples) were obtained from 21 farms distributed in four geographical regions in Colombia. Salmonella spp. microbiological and molecular detection were determined by two Salmonella spp. MDS3M™ and MALDI-TOF MS assays, respectively. In addition, for serological evaluation 231 serum samples were analyzed employing ELISA Salmonella Pigtype®-Salmonella Ab (QUIAGEN®). Additionally, 41 isolates were tested for antimicrobial susceptibility using broth microdilution technique (Panel B1016-180 Beckman Coulter NC72®) and verified with WHONET 2016 software. Risk factors were assessed from a survey and analyzed for statistical significance by U Mann-Whitney test. An 8.9% prevalence (n=45) and 38.1% (n=88) seroprevalence were determined. All isolates presented 100% antimicrobial susceptibility against amikacin. However, resistance against penicillin, tetracycline, cefuroxime and trimethoprim/sulfamethoxazole was present in more than 50% of evaluated strains. Risk factors associated with Salmonella spp. presence were surface water use, rough-surfaced on floors, presence of hoppers as feeders and worker's boots. Bacteria were present in animals and environmental samples from evaluated farms. Animal contact and/or exposure with the microorganism were also evident in obtained serological response. Bacteria presence depended on management practices and infrastructure, likewise antibiotic use, supplemented in the diet may have induced an increase in Salmonella spp. antimicrobial resistance.(AU)

Para determinar Salmonellaspp. prevalência/soroprevalência, padrões de resistência antimicrobiana e identificação de fatores de risco associados à sua presença em granjas suínas colombianas. Foram obtidas 504 amostras (fezes, zaragatoas e amostras do ambiente) de 21 fazendas distribuídas em quatro regiões geográficas da Colômbia. Salmonella spp., a detecção microbiológica e molecular foi determinada por 2 Salmonella spp. Ensaios MDS3M™ e MALDI-TOF MS, respectivamente. Além disso, para avaliação sorológica, foram analisadas 231 amostras de soro empregando ELISA Salmonella Pigtype® - Salmonella Ab (QUIAGEN®). Além disso, 41 isolados foram testados quanto à suscetibilidade antimicrobiana usando a técnica de microdiluição em caldo (Painel B1016-180 Beckman Coulter NC72®) e verificados com o software WHONET 2016. Os fatores de risco foram avaliados em uma pesquisa e analisados quanto à significância estatística pelo teste U Mann-Whitney. Foram determinadas prevalências de 8,9% (n=45) e 38,1% (n=88). Todos os isolados apresentaram 100% de suscetibilidade antimicrobiana à amicacina. No entanto, resistência à penicilina, tetraciclina, cefuroxima e trimetoprim/sulfametoxazol estava presente em mais de 50% das cepas avaliadas. Fatores de risco associados à Salmonella spp., presença de uso de água de superfície, superfície áspera no chão, presença de tremonhas como alimentadores e botas de trabalho. Bactérias estavam presentes em animais e amostras ambientais de fazendas avaliadas. O contato animal e/ou a exposição ao microrganismo também foram evidentes na resposta sorológica obtida. A presença de bactérias dependia de práticas de manejo e infraestrutura, assim como o uso de antibióticos suplementados na dieta pode ter induzido um aumento de Salmonella spp. resistência antimicrobiana.(AU)

Towards a subunit vaccine from a Shigella flexneri ΔtolR mutant.

Disruption of one or more components of the Tol-Pal system, involved in maintaining the integrity of the outer membrane of Gram-negative bacteria, has been proposed as a method to increase the yield obtained from natural production of outer membrane vesicles (OMV). We present a new OMV-based product, obtained from genetically modified Shigella flexneri 2a with a non-polar deletion in tolR and heat-inactivated (HT-ΔtolR). The S. flexneri ΔtolR strain lead to a higher release of vesicles, more than 8-times when compared to the yield obtained from chemically inactivated wild type strain. S. flexneri mutant strain appeared to be more sensitive to different chemical compounds, including antibiotics, bile salts or human complement and it was also less virulent in both in vitro and in vivo assays. The mutation produced some changes in the LPS O-chain and protein expression. S. flexneri ΔtolR was enriched in long and very long LPS O-chain and expressed a different pattern of surface proteins or lipoproteins. In vitro toxicity and activation properties were determined in Raw 267.4 macrophage cell line. HT-ΔtolR antigenic complex was non-cytotoxic and activation markers, such as MHC-II or CD40, were highly expressed during incubation with this product. Finally, preliminary studies on the antibody response elicited by HT-ΔtolR demonstrated a robust and diverse response in mice. Considering these promising results, HT-ΔtolR antigenic extract appears as a new potential vaccine candidate to face shigellosis.

Analysis of Favorable Process Conditions for the Manufacturing of Thin-Wall Pieces of Mild Steel Obtained by Wire and Arc Additive Manufacturing (WAAM).

One of the challenges in additive manufacturing (AM) of metallic materials is to obtain workpieces free of defects with excellent physical, mechanical, and metallurgical properties. In wire and arc additive manufacturing (WAAM) the influences of process conditions on thermal history, microstructure and resultant mechanical and surface properties of parts must be analyzed. In this work, 3D metallic parts of mild steel wire (American Welding Society-AWS ER70S-6) are built with a WAAM process by depositing layers of material on a substrate of a S235 JR steel sheet of 3 mm thickness under different process conditions, using as welding process the gas metal arc welding (GMAW) with cold metal transfer (CMT) technology, combined with a positioning system such as a computer numerical controlled (CNC) milling machine. Considering the hardness profiles, the estimated ultimate tensile strengths (UTS) derived from the hardness measurements and the microstructure findings, it can be concluded that the most favorable process conditions are the ones provided by CMT, with homogeneous hardness profiles, good mechanical strengths in accordance to conditions defined by standard, and without formation of a decohesionated external layer; CMT Continuous is the optimal option as the mechanical properties are better than single CMT.

Prevalence of Salmonella spp., in mesenteric pig's ganglia at Colombian benefit plants / Prevalencia de Salmonella spp., en ganglios mesentéricos de porcinos en plantas de beneficio Colombianas

ABSTRACT Objective. To determine the prevalence of Salmonella spp., in pigs mesenteric ganglion, from different regions of Colombia. Materials and Methods. A stratified sampling by proportional fixation was carried out at benefit plants of each of the 13 participating departments, whose pork production volume is representative at national level. Sampling was performed during five months, for a total of 457 samples analyzed. Salmonella spp., identification was performed by the MDS Molecular System, later isolates were confirmed in Maldi-TOF MS. Antimicrobial susceptibility of the isolates was determined using the B1016-180 panel and statistical analysis was performed in Whonet 2016, some of the multi-resistant isolates were them serotyped by Kauffman-White method. Results. National prevalence was 28.2%, with the presence of S. Typhimurium, S. Agama, S. London, S. Agona, S. Haifa and S. 1,4,12: i: -. Resistance to antibiotics frequently used in human (23.6% Trimethoprim/Sulfamethoxazole, 2.7% Cefotaxime (CTX), 11.8% Ampicillin (AMP) and 1.8% Ciprofloxacin) was found. Conclusion. The prevalence of Salmonella in mesenteric ganglia was 28.2%, being the Huila region the one with the highest prevalence, recovering atypical serotypes such as S. London and S. Haifa.

RESUMEN Objetivo. Determinar la prevalencia de Salmonella spp., en ganglios mesentéricos de porcinos, provenientes de diferentes regiones de Colombia. Materiales y Métodos. Se realizó un muestreo estratificado por fijación proporcional en plantas de beneficio, de cada uno de los 13 departamentos participantes, cuyo volumen de producción de carne de cerdo es representativo a nivel nacional. El muestreo se realizó durante cinco meses, para un total de 457 muestras analizadas. La identificación de Salmonella spp., se realizó mediante el Sistema Molecular MDS, luego los aislamientos fueron confirmados por Maldi-TOF MS. Se determinó la susceptibilidad antimicrobiana de los aislamientos usando el panel B1016-180 y el análisis estadístico se realizó en Whonet 2016, posteriormente algunos de los aisalmientos multi-resistentes fueron serotipificados por el método de Kauffman-White. Resultados. La prevalencia nacional fue 28.2%, con presencia de los serotipos S. Typhimurium, S. Agama, S. London, S. Agona, S. Haifa y S. 1,4,12: i : --. Se encontró resistencia a antibióticos de uso frecuente en humanos (23.6% Trimetoprim/Sulfametoxazol, 2.7% Cefotaxime (CTX), 11.8% Ampicilina (AMP) y 1.8% Ciprofloxacina). Conclusión. La prevalencia de Salmonella en ganglios mesénticos fue del 28.2%, siendo la región del Huila la que más aportó, se recuperaron serotipos atípicos como S. London y S. Haifa.

Detección e identificación de Salmonella spp. en huevos para consumo humano, provenientes de diferentes localidades de Bogotá, Colombia, 2015 / Salmonella spp. isolation and identification in eggs for human consumption from different urban areas in Bogotá, Colombia, 2015

Objetivo: Determinar la prevalencia de Salmonella spp. en muestras de huevos para consumo humano en localidades de Bogotá. Materiales y métodos: Se obtuvieron 96 muestras en tiendas y plazas de mercado de 4 localidades de la ciudad. Se procesó de forma separada la cáscara y el contenido interno mediante métodos microbiológicos y moleculares para aislamiento e identificación Salmonella spp. Resultados: Se determinó una prevalencia total de Salmonella spp. de 9,4% (n=9), de ésta el 55% (n = 5) provenían del contenido interno y 44% (n = 4) de cáscara, sin embargo no se logró tipificar el serovar de Salmonella enterica presente. Las localidades con mayor presencia del patógeno fueron Usaquén y Fontibón. Discusión: Estudios realizados en Colombia evidencian bajas prevalencias de Salmonella spp. (0 -1,74%) en muestras de huevos, sin embargo los hallazgos de este estudio evidencian una mayor recuperación, lo que podría asociarse con inadecuadas condiciones de manejo y/o almacenamiento del producto. Conclusión: Se estableció la presencia de Salmonella spp. en las muestras de huevo evaluadas, lo que implica un riesgo potencial para la salud pública, por lo que es necesario ampliar este tipo de estudios para conocer la situación real a nivel nacional frente a este patógeno.

Objective: To determine the prevalence of Salmonella spp presence in eggs for human consumption in urban areas in Bogota. Materials and methods: 96 samples were collected from convenience stores and markets in 4 urban areas in the city. The eggshells were separated from the egg's internal content and both were processed separately using microbiological and molecular techniques to isolate and identify Salmonella spp. strains. Results: A Salmonella spp. prevalence of 9.4% (n=9) was found. Salmonella spp was isolated from the egg's internal content in 55% (n=5) of samples and 44% (n=4) from the eggshells. The Salmonella enterica serovar could not be identified. The pathogen was more frequently isolate in samples from Usaquén and Fontibón urban areas. Discussion: Studies of Salmonella spp. prevalence in eggs done in Colombia have shown it to be low (0-1.74%); However, this study determined a higher prevalence. These results suggest that inadequate handling/storage conditions could have been associated with them. Conclusion: Salmonella spp. was isolated from the egg samples from 4 different urban areas in Bogotá. These findings suggest the existence of a public health risk; therefore, there is a need to perform wider and more complete studies to determine the actual situation of Salmonella ssp. egg contamination in the country.

New ATP8A2 gene mutations associated with a novel syndrome: encephalopathy, intellectual disability, severe hypotonia, chorea and optic atrophy.

We report the clinical and biochemical findings from two unrelated patients who presented with a novel syndrome: encephalopathy, intellectual disability, severe hypotonia, chorea and optic atrophy. Whole exome sequencing (WES) uncovered a homozygous mutation in the ATP8A2 gene (NM_016529:c.1287G > T, p.K429N) in one patient and compound heterozygous mutations (c.1630G > C, p.A544P and c.1873C > T, p.R625W) in the other. Only one haploinsufficiency case and a family with a homozygous mutation in ATP8A2 gene (c.1128C > G, p.I376M) have been described so far, with phenotypes that differed slightly from the patients described herein. In conclusion, our data expand both the genetic and phenotypic spectrum associated with ATP8A2 gene mutations.

Clinical, neuroradiological and molecular characterization of cerebellar dysplasia with cysts (Poretti-Boltshauser syndrome).

Cerebellar dysplasia with cysts and abnormal shape of the fourth ventricle, in the absence of significant supratentorial anomalies and of muscular involvement, defines recessively inherited Poretti-Boltshauser syndrome (PBS). Clinical features comprise non-progressive cerebellar ataxia, intellectual disability of variable degree, language impairment, ocular motor apraxia and frequent occurrence of myopia or retinopathy. Recently, loss-of-function variants in the LAMA1 gene were identified in six probands with PBS. Here we report the detailed clinical, neuroimaging and genetic characterization of 18 PBS patients from 15 unrelated families. Biallelic LAMA1 variants were identified in 14 families (93%). The only non-mutated proband presented atypical clinical and neuroimaging features, challenging the diagnosis of PBS. Sixteen distinct variants were identified, which were all novel. In particular, the frameshift variant c.[2935delA] recurred in six unrelated families on a shared haplotype, suggesting a founder effect. No LAMA1 variants could be detected in 27 probands with different cerebellar dysplasias or non-progressive cerebellar ataxia, confirming the strong correlate between LAMA1 variants and PBS.

Práctica clínica cotidiana frente a la evidencia científica en el manejo fisioterapéutico del dolor lumbar crónico inespecífico / Daily Clinical Practice of Scientific Evidence in the Physiotherapeutic Management of Chronic Nonspecific Low Back Pain / Prática clínica cotidiana versus a evidência científica no manejo fisioterapêutico da dor lombar crónica inespecífica

Introducción: El 80% de los adultos experimenta dolor lumbar por lo menos una vez en su vida. El dolor lumbar es la tercera causa de consulta en urgencias, la cuarta en medicina general, la segunda de pensión por invalidez y la primera de reubicación laboral. Objetivo: Comparar los criterios que orientan la toma de decisiones de un grupo de fisioterapeutas en Bogotá, Colombia para el manejo del dolor lumbar crónico inespecífico con los criterios de manejo contenidos en la guía COST B13 (European Guidelines for the Management of Chronic Non-Specific Low Back Pain, 2004). Materiales y método: Se trató de un estudio descriptivo, en el cual se seleccionó la guía de práctica clínica COST B13 para el manejo de dolor lumbar crónico inespecífico mediante la herramienta AGREE y se aplicó una encuesta a 50 fisioterapeutas por medio de un muestreo por conveniencia con el fin de comparar las prácticas clínicas que se realizan frente a las recomendaciones dadas en la guía. Resultados: El 56% de los encuestados contaba con algún tipo de entrenamiento para el manejo del dolor lumbar crónico inespecífico (DLCI). El 94% de los pacientes con DLCI atendidos oscilan en edades de 40 a 59 años, con predominio del sexo femenino. El 80% de los fisioterapeutas encuestados manifestó que la ayuda diagnóstica con la que cuentan para el manejo de los pacientes es la imagen radiológica. El 80% de los fisioterapeutas evalúa la variable de dolor lumbar experimentado por el paciente y el 54% la postura. Otros aspectos fueron reportados en menor porcentaje como test neurales, rangos de movimiento y pruebas de flexibilidad muscular. En el tratamiento del DLCI, los fisioterapeutas reportan hacer uso de los estiramientos en el 80% de los casos, la termoterapia superficial, en el 70% y el fortalecimiento muscular isométrico, en el 70%, todos con resultados favorables. Conclusión: Existen diferencias entre la práctica clínica y los lineamientos contenidos en las recomendaciones dadas por la guía COST B13 para DLCI, principalmente en los procesos de evaluación clínica, ya que suelen estar centrados en la observación y no siempre en la medición rigurosa, lo que dificulta la posibilidad de establecer indicadores de proceso y desenlace en el diagnóstico y tratamiento de los pacientes con DLCI.

Introduction: 80% of adults experience back pain at least once in their life.Back pain is the third leading cause of consultation in the emergency room, the fourth in general practice, the second of disability pension and the first job relocation. Objective: To compare criteria that guide decision making of a group of physiotherapists in Bogota, Colombia, for managing chronic nonspecific low back pain management criteria contained in the guide COST B13 (European Guidelines for the Management of Chronic Non-Specific Low Back Pain, 2004). Material and methods: This was a descriptive study, for which the clinical practice guideline COST B13 for the management of chronic nonspecific low back pain through the AGREE tool is selected. A survey was applied to 50 physiotherapists through a convenience sample to compare the clinical practices performed with the recommendations given guidance. Results: 56% of respondents had some type of training for the management of chronic nonspecific low back pain (cnlbp). 94% of attended patients with cnlbp ranged ages from 40 to 59, with female predominance. 80% of respondents stated that physiotherapists' diagnostic help with counting for the management of patients is the radiological image. 80 % of physiotherapists evaluate the variable of experienced lumbar pain based on what the patient says and 54% based on the posture. Other aspects reported rated lower percentages. In the treatment of cnlbp, physiotherapists reported the use of stretching in 80% of cases; superficial thermotherapy in 70% and isometric muscle strength in 70% of cases, all with favorable results. Conclusion: There are differences between the physiotherapists' clinical practice of and the guidelines contained in the recommendations of the in the cnlbp cost B13 guide. Mainly in the processes of physiotherapeutic assessment, as they are often focused on observation and not always in the rigorous measurement, which makes it difficult to establish process indicators and results in the diagnosis and treatment of patients with cnlbp.

Introdução: o 80% dos adultos experimenta dor lombar pelo menos uma vez em sua vida; a dor lombar é a terceira causa de consulta em emergências, a quarta em medicina geral, a segunda de pensão por invalidez e a primeira de deslocalização laboral. Objetivo: Comparar os critérios que orientam a tomada de decisões de um grupo de fisioterapeutas em Bogotá, Colômbia, para o manejo da dor lombar crónica inespecífico com os critérios de manejo conteúdos na guia COST B13 (European Guidelines for the Management of Chronic Non-Specific Low Back Pain, 2004). Materiais e métodos: tratou-se de um estudo descritivo, no qual se selecionou a guia de prática clínica COSTB 13 para o manejo de dor lombar crónico inespecífico através da ferramenta AGREE e se aplicou uma enquete a 50 fisioterapeutas através de uma amostragem por conveniência com o fim de comparar as práticas clínicas que se realizam frente às recomendações dadas na guia. Resultados: o 56% dos questionados contavam com algum tipo de treinamento para o manejo da dor lombar crónico inespecífico (DLCI). O 94% dos pacientes com DLCI atendidos oscilam em idades de 40 a 59 anos, com predomínio do sexo feminino. O 80% dos fisioterapeutas avalia a variável de dor lombar experimentado pelo paciente e o 54% a postura. Outros aspectos foram reportados em menor porcentagem como teste neurais, rasgos de movimentos, e provas de flexibilidade muscular. No tratamento da DLCI, os fisioterapeutas reportam fazer uso dos estiramentos no 80% dos casos, a termoterapia superficial no 70% e o fortalecimento muscular isométrico no 70%, todos com resultados favoráveis. Conclusões: Existem diferenças entre a prática clínica e os lineamentos conteúdos nas recomendações dadas pela guia COST B12 para DLCI principalmente nos processos de avaliação clínica devido a que costumam estar centrados na observação e não sempre na medição rigorosa o que dificulta a possibilidade de estabelecer indicadores de processo e desenlace no diagnóstico e tratamento dos pacientes com DLCI.

Immunogenicity of peanut proteins containing poly(anhydride) nanoparticles.

In the last decade, peanut allergy has increased substantially. Significant differences in the prevalence among different countries are attributed to the type of thermal processing. In spite of the high prevalence and the severe reaction induced by peanuts, there is no immunotherapy available. The aim of this work was to evaluate the potential application of poly(anhydride) nanoparticles (NPs) as immunoadjuvants for peanut oral immunotherapy. NPs loaded with raw or roasted peanut proteins were prepared by a solvent displacement method and dried by either lyophilization or spray-drying. After physicochemical characterization, their adjuvant capacity was evaluated after oral immunization of C57BL/6 mice. All nanoparticle formulations induced a balanced T(H)1 and T(H)2 antibody response, accompanied by low specific IgE induction. In addition, oral immunization with spray-dried NPs loaded with peanut proteins was associated with a significant decrease in splenic T(H)2 cytokines (interleukin 4 [IL-4], IL-5, and IL-6) and enhancement of both T(H)1 (gamma interferon [IFN-γ]) and regulatory (IL-10) cytokines. In conclusion, oral immunization with poly(anhydride) NPs, particularly spray-dried formulations, led to a pro-T(H)1 immune response.

Consensus design of a NOD receptor leucine rich repeat domain with binding affinity for a muramyl dipeptide, a bacterial cell wall fragment.

Repeat proteins have recently emerged as especially well-suited alternative binding scaffolds due to their modular architecture and biophysical properties. Here we present the design of a scaffold based on the consensus sequence of the leucine rich repeat (LRR) domain of the NOD family of cytoplasmic innate immune system receptors. Consensus sequence design has emerged as a protein design tool to create de novo proteins that capture sequence-structure relationships and interactions present in nature. The multiple sequence alignment of 311 individual LRRs, which are the putative ligand-recognition domain in NOD proteins, resulted in a consensus sequence protein containing two internal and N- and C-capping repeats named CLRR2. CLRR2 protein is a stable, monomeric, and cysteine free scaffold that without any affinity maturation displays micromolar binding to muramyl dipeptide, a bacterial cell wall fragment. To our knowledge, this is the first report of direct interaction of a NOD LRR with a physiologically relevant ligand.