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Exome and genome sequencing (ES/GS) are routinely used for the diagnosis of genetic diseases in developed countries. However, their implementation is limited in countries from Latin America. We aimed to describe the results of GS in patients with suspected rare genetic diseases in Colombia. We studied 501 patients from 22 healthcare sites from January to December 2022. GS was performed in the index cases using dried blood spots on filtercards. Ancestry analysis was performed under iAdmix. Multiomic testing was performed when needed (biomarker, enzymatic activity, RNA-seq). All tests were performed at an accredited genetic laboratory. Ethnicity prediction data confirmed that 401 patients (80%) were mainly of Amerindian origin. A genetic diagnosis was established for 142 patients with a 28.3% diagnostic yield. The highest diagnostic yield was achieved for pathologies with a metabolic component and syndromic disorders (p < 0.001). Young children had a median of 1 year of diagnostic odyssey, while the median time for adults was significantly longer (15 years). Patients with genetic syndromes have spent more than 75% of their life without a diagnosis, while for patients with neurologic and neuromuscular diseases, the time of the diagnostic odyssey tended to decrease with age. Previous testing, specifically karyotyping or chromosomal microarray were significantly associated with a longer time to reach a definitive diagnosis (p < 0.01). Furthermore, one out of five patients that had an ES before could be diagnosed by GS. The Colombian genome project is the first Latin American study reporting the experience of systematic use of diagnostic GS in rare diseases.
Assuntos
Doenças Raras , Sequenciamento Completo do Genoma , Humanos , Colômbia , Doenças Raras/genética , Doenças Raras/diagnóstico , Adulto , Masculino , Feminino , Criança , Sequenciamento Completo do Genoma/normas , Adolescente , Pré-Escolar , Testes Genéticos/métodos , Testes Genéticos/normas , Pessoa de Meia-Idade , LactenteRESUMO
Molecular diagnostic tests have evolved very rapidly in the field of human health, especially with the arrival of the recent pandemic caused by the SARS-CoV-2 virus. However, the animal sector is constantly neglected, even though accurate detection by molecular tools could represent economic advantages by preventing the spread of viruses. In this regard, the swine industry is of great interest. The main viruses that affect the swine industry are described in this review, including African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), and porcine circovirus (PCV), which have been effectively detected by different molecular tools in recent times. Here, we describe the rationale of molecular techniques such as multiplex PCR, isothermal methods (LAMP, NASBA, RPA, and PSR) and novel methods such as CRISPR-Cas and microfluidics platforms. Successful molecular diagnostic developments are presented by highlighting their most important findings. Finally, we describe the barriers that hinder the large-scale development of affordable, accessible, rapid, and easy-to-use molecular diagnostic tests. The evolution of diagnostic techniques is critical to prevent the spread of viruses and the development of viral reservoirs in the swine industry that impact the possible development of future pandemics and the world economy.
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Resumen Introducción: la ventilación mecánica es una práctica común en las unidades de cuidados intensivos y anestesiología. Tiene implicaciones terapéuticas, pero también es potencialmente nociva para el sistema respiratorio y los órganos distantes, por lo cual es imprescindible monitorizar los parámetros ventilatorios de manera continua. Objetivo: describir la ecuación del movimiento del sistema respiratorio y sus aplicaciones clínicas en el paciente bajo ventilación mecánica. Desarrollo: la ecuación del movimiento del sistema respiratorio integra las fuerzas dinámicas generadas por el ventilador y las propiedades intrínsecas del pulmón y la caja torácica. Expresa la presión en el sistema respiratorio en relación con el volumen, la elastancia, la resistencia, el flujo de aire y las presiones generadas por el ventilador y el paciente. Las presiones elevadas en el sistema respiratorio se asocian a una mayor mortalidad en pacientes con ventilación mecánica, por lo que la identificación de los componentes que condicionan la elevación de las presiones mediante la ecuación del movimiento del sistema respiratorio permite realizar modificaciones a los parámetros programados del ventilador para mantener una ventilación protectora. Conclusión: la toma de decisiones basada en la ecuación del movimiento del sistema respiratorio permite realizar ajustes en los parámetros ventilatorios según las características y enfermedades del paciente bajo ventilación mecánica.
Abstract Introduction: Mechanical ventilation is a common practice in intensive care units and anesthesiology with both therapeutic and potentially harmful implications for the respiratory system and distant organs, that is why it is of utmost importance to continually monitor ventilation parameters. Objective: To describe the equation of motion of the respiratory system and its clinical applications in the patient under invasive mechanical ventilation. Main: The equation of motion of the respiratory system integrates the dynamic forces generated by the ventilator with the intrinsic properties of the lung and chest wall. It expresses the pressure in the respiratory system in relation to volume, elastance, resistance, air flow and pressures generated by the ventilator and the patient. Elevated pressures in the respiratory system during mechanical ventilation are associated with greater mortality, that is why the identification of the components responsible for elevation of pressures through the equation of motion of the respiratory system allows to modify ventilator programmed parameters to maintain a protective ventilation. Conclusion: Decision-making based on the equation of motion of the respiratory system allows to modify ventilatory parameters according to the characteristics and diseases of the patient under mechanical ventilation.
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Feruloyl esterases (FAEs) are versatile enzymes able to release hydroxycinnamic acids or synthesize their ester derivatives, both molecules with interesting biological activities such as: antioxidants, antifungals, antivirals, antifibrotic, anti-inflammatory, among others. The importance of these molecules in medicine, food or cosmetic industries provides FAEs with several biotechnological applications as key industrial biocatalysts. However, FAEs have some operational limitations that must be overcome, which can be addressed through different protein engineering approaches to enhance their thermal stability, catalytic efficiencies, and selectivity. This review aims to present a brief historical tour through the mutagenesis strategies employed to improve enzymes performance and analyze the current protein engineering strategies applied to FAEs as interesting biocatalysts. Finally, an outlook of the future of FAEs protein engineering approaches to achieve successful industrial biocatalysts is given.
Assuntos
Hidrolases de Éster Carboxílico , Engenharia de Proteínas , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Ácidos Cumáricos/metabolismo , Biotecnologia , Catálise , Biocatálise , Enzimas/metabolismoRESUMO
Halophilic microorganisms are potentially capable as platforms to produce low-cost biosurfactants. However, the robustness of bioprocesses is still a challenge and, therefore, it is essential to understand the effects of microbiological culture conditions through bioreactor engineering. Based on a design of experiments (DOE) and a response surface methodology (RSM) tailored and taken from the literature, the present work focuses on the evaluation of a composite central design (CCD) under batch cultures in stirred-tank bioreactors with the halophilic bacteria Salibacterium sp. 4CTb in order to determine the operative conditions that favor mass transfer and optimize the production of a lipopeptide. The results obtained showed profiles highlighting the most favorable culture conditions, which lead to an emulsification index (E24%) higher than 70%. Moreover, through the behavior of dissolved oxygen (DO), it was possible to experimentally evaluate the higher volumetric coefficient of mass transfer in the presence of lipopeptide (kLa = 31 1/h) as a key criterion for the synthesis of the biosurfactant on further cell expansion.
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Reportamos el caso de una adolescente femenina que ingresa al Hospital Mario Catarino Rivas en San Pe- dro Sula con antecedente de tricofagia desde hace 5 años. Ingresa con historia de dolor abdominal epigás- trico de 3 meses de evolución con náuseas y vómitos de alimentos ocasionales junto a pérdida de aproxima- damente 6 kilogramos de peso en los últimos 3 meses. Paciente fue ingresada y se completaron estudios diagnosticándose tricobezoar gástrico con poste- rior intervención quirúrgica: laparotomía media supraumbilical accediendo al estómago sobre el cual se le realizó una gastrostomía antral de 6 cm de longitud por donde se extrae el bezoar en su to- talidad. Se cierra posteriormente la gastrotomía.
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Humanos , Feminino , Criança , Tricotilomania , Bezoares/diagnósticoRESUMO
El síndrome de Nicolau o Embolia cutis medicamentosa o Dermatitis livedoide, es un síndrome raro en pediatría que se presenta como complicación de una inyección intramuscular con múltiples grados de daño tisular, in- cluyendo la necrosis de la piel y de los tejidos profundos. Reportamos el caso de un paciente masculino de 2 años de edad, sin antecedentes patológicos que acudió a la emergencia de pediatría del Hospital "Mario Catarino Rivas" (HMCR) en San Pedro Sula, Cortes, con historia de presentar lesión necrótica en la extremidad inferior y glúteo izquierdo, dolorosa a la palpación, que apa- reció posterior a la administración de un medicamen- to compuesto por vía intramuscular. Al examen físico se encontró una extensa placa necrótica dolorosa a la palpación y a la movilización del miembro, con pulsos periféricos conservados y sin frialdad distal. A través de abordaje multidisciplinario se brindó limpieza qui- rúrgica, cobertura antibiótica, manejo antitrombótico e injerto, con evolución favorable, por lo que resalta- mos que la identificación y manejo multidisciplinario oportuno es fundamental, como lo fue en este caso...(AU)
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Humanos , Masculino , Pré-Escolar , Penicilina G Benzatina , Síndrome de Nicolau/diagnóstico , Injeções Intramusculares/métodos , Necrose/complicaçõesRESUMO
INTRODUCTION: Simultaneous mechanical ventilation of several patients with a single ventilator might reduce the deficit of these devices for the care of patients with acute respiratory failure due to Covid-19. OBJECTIVE: To communicate the results of a mechanical ventilation exercise with a ventilator in a lung simulator, and simultaneously in two and four. RESULTS: No statistically significant differences were observed between programmed, recorded and measured positive end-expiratory pressure, mean airway pressure and peak pressure, except when simultaneously ventilating four lung simulators. CONCLUSIONS: Simultaneous mechanical ventilation should be implemented by medical personnel with experience in the procedure, be restricted to two patients and carried out in the intensive care unit.
INTRODUCCIÓN: La ventilación mecánica simultánea a varios pacientes con un solo ventilador podría disminuir el déficit de esos dispositivos para atender a los enfermos con insuficiencia respiratoria aguda por Covid-19. OBJETIVO: Comunicar los resultados de un ejercicio de ventilación mecánica con un ventilador en un simulador de pulmón, y simultáneamente en dos y cuatro. RESULTADOS: No se observaron diferencias estadísticamente significativas entre la presión positiva al final de la espiración, presión media de la vía aérea y presión pico programadas, registradas y medidas, excepto al ventilar simultáneamente cuatro simuladores de pulmón. CONCLUSIONES: La ventilación mecánica simultánea debe ser instaurada por personal médico con experiencia en el procedimiento, restringirse a dos pacientes y ser realizada en la unidad de cuidados intensivos.
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Infecções por Coronavirus/terapia , Pneumonia Viral/terapia , Respiração Artificial/métodos , Ventiladores Mecânicos/provisão & distribuição , COVID-19 , Infecções por Coronavirus/fisiopatologia , Desenho de Equipamento , Humanos , Unidades de Terapia Intensiva , Pandemias , Pneumonia Viral/fisiopatologia , Respiração com Pressão Positiva , Respiração Artificial/instrumentação , Insuficiência Respiratória/terapia , Insuficiência Respiratória/virologiaRESUMO
Resumen Introducción: La ventilación mecánica simultánea a varios pacientes con un solo ventilador podría disminuir el déficit de esos dispositivos para atender a los enfermos con insuficiencia respiratoria aguda por Covid-19. Objetivo: Comunicar los resultados de un ejercicio de ventilación mecánica con un ventilador en un simulador de pulmón, y simultáneamente en dos y cuatro simuladores. Resultados: No se observaron diferencias estadísticamente significativas entre la presión positiva al final de la espiración, presión media de la vía aérea y presión pico programadas, registradas y medidas, excepto al ventilar simultáneamente cuatro simuladores de pulmón. Conclusiones: La ventilación mecánica simultánea debe ser instaurada por personal médico con experiencia en el procedimiento, restringirse a dos pacientes y ser realizada en la unidad de cuidados intensivos.
Abstract Introduction: Simultaneous mechanical ventilation of several patients with a single ventilator might reduce the deficit of these devices for the care of patients with acute respiratory failure due to Covid-19. Objective: To communicate the results of a mechanical ventilation exercise with a ventilator in a lung simulator, and simultaneously in two and four. Results: No statistically significant differences were observed between positive end-expiratory pressure, mean airway pressure, and programmed, recorded and measured peak pressure, except when simultaneously ventilating four lung simulators. Conclusions: Simultaneous mechanical ventilation should be implemented by medical personnel with experience in the procedure, be restricted to two patients and carried out in the intensive care unit.
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Humanos , Pneumonia Viral/terapia , Respiração Artificial/métodos , Ventiladores Mecânicos/provisão & distribuição , Infecções por Coronavirus/terapia , Pneumonia Viral/fisiopatologia , Respiração Artificial/instrumentação , Insuficiência Respiratória/terapia , Insuficiência Respiratória/virologia , Respiração com Pressão Positiva , Infecções por Coronavirus/fisiopatologia , Desenho de Equipamento , Pandemias , COVID-19 , Unidades de Terapia IntensivaRESUMO
Breast cancer is a worldwide health problem, and the complexity of the disease, as well as the lack of treatment specificity, generates an urgent need for developing prophylactic and therapeutic measures. Searching for novel epitope-based approaches able to induce tumour immunity, we designed virus-like particles (VLPs) derived from Human parvovirus B19 assembled of chimeric VP2 proteins displaying two epitopes from the insulin-like growth factor-1 receptor (IGF-1R). Here, we present the generation of two chimeric VP2s that retain the stability, solubility and conditions of purification and assembly of the native VP2. We generated versatile chimeric multiepitope anti-cancer vaccine candidates, which prevented and delayed tumour growth when used in a prophylactic scheme of 4 weekly immunizations prior to 4T1 cell inoculation in female BALB/c mice. The presence of specific antibodies against the displayed epitopes suggests their participation in the protective effect; in contrast, no significant proliferative T-cell responses were recorded following stimulation by specific epitopes. The results comprise an approach whereby fusing desired epitopes from cancer to the N-terminus of B19 VP2 protein can generate a library of chimeric VP2-desired epitopes for further assembly in a designed and personalized epitope delivery system.
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Neoplasias da Mama/prevenção & controle , Epitopos/metabolismo , Parvovirus B19 Humano/metabolismo , Receptor IGF Tipo 1/imunologia , Vacinas de Partículas Semelhantes a Vírus/administração & dosagem , Animais , Apoptose , Neoplasias da Mama/imunologia , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/imunologia , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular , Epitopos/genética , Feminino , Humanos , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Parvovirus B19 Humano/genética , Resultado do Tratamento , Vacinas de Partículas Semelhantes a Vírus/genética , Vacinas de Partículas Semelhantes a Vírus/imunologiaRESUMO
The evaluation of first-degree family members is very important to detect additional cases of polyglandular autoimmune syndrome type 2. The genetic evaluation of human leukocyte antigen (HLA) may be useful in the study of this syndrome. This study is the first report of an HLA study of this syndrome in a Mexican family.
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Background: Aspergillus ochraceus was isolated from coffee pulp and selected as an interesting hydroxycinnamoyl esterase strain producer, using an activity microplate high-throughput screening method. In this work, we purified and characterized a new type C A. ochraceus feruloyl esterase (AocFaeC), which synthesized specifically butyl hydroxycinnamates in a ternary solvent system. Results: AocFaeC was produced by solid state fermentation, reaching its maximal activity (1.1 U/g) after 48 h of culture. After purification, the monomeric protein (34 kDa) showed a specific activity of 57.9 U/mg towards methyl ferulate. AocFaeC biochemical characterization confirmed its identity as a type C feruloyl esterase and suggested the presence of a catalytic serine in the active site. Its maximum hydrolytic activity was achieved at 40°C and pH 6.5 and increased by 109 and 77% with Ca2+ and Mg2+, but decreased by 90 and 45% with Hg2+ and Cu2+, respectively. The initial butyl ferulate synthesis rate increased from 0.8 to 23.7 nmol/min after transesterification condition improvement, using an isooctane:butanol:water ternary solvent system, surprisingly the synthesis activity using other alcohols was negligible. At these conditions, the synthesis specific activities for butyl p-coumarate, sinapinate, ferulate, and caffeate were 87.3, 97.6, 168.2, and 234 U/µmol, respectively. Remarkably, AocFaeC showed 5 folds higher butyl caffeate synthesis rate compared to type B Aspergillus niger feruloyl esterase, a well-known enzyme for its elevated activity towards caffeic acid esters. Conclusions: Type C feruloyl esterase from A. ochraceus is a butanol specific biocatalyst for the synthesis of hydroxycinnamates in a ternary solvent system
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Aspergillus ochraceus/enzimologia , Hidrolases de Éster Carboxílico/metabolismo , Ácidos Cumáricos/síntese química , Solventes , Espectrofotometria , Hidrolases de Éster Carboxílico/isolamento & purificação , Cromatografia , Café , Butanóis , Eletroforese , FermentaçãoRESUMO
Solid-state fermentation (SSF) has been largely employed during the last three decades to produce different biomolecules of industrial interest, particularly enzymes. Through the use of agroindustrial wastes as SSF substrates, an economic process of lipases production can be achieved. In this chapter we describe a comprehensive SSF method for producing an economical preparation of Rhizomucor miehei lipase, employing sugarcane bagasse and used vegetal oil as substrates. To demonstrate the usefulness of the lipase produced by this method, we utilized directly the dried fermented solid, as a heterogeneous biocatalyst for the ethanolysis of different fats and oils. Final ethyl ester conversions (>90%, 24 h) were similar with those obtained using a commercial immobilized Rhizomucor miehei lipase at our best conditions. In this work we demonstrated that SSF is an easy and economical method for the production of lipases that can be used directly as heterogeneous biocatalysts for biodiesel production, employing low-cost feedstocks.
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Bioengenharia , Fermentação , Lipase/biossíntese , Bioengenharia/instrumentação , Bioengenharia/métodos , Biocombustíveis , Catálise , Concentração de Íons de Hidrogênio , Hidrólise , Cinese , Lipase/isolamento & purificação , TemperaturaRESUMO
Resumen: La mayoría de los pacientes que requieren ventilación > 24 horas y mejoran pueden ser extubados tras la primera prueba de ventilación espontánea. El reto es mejorar la desconexión de los pacientes que fracasan. El retiro de la ventilación mecánica es un elemento esencial en el cuidado de los pacientes críticamente enfermos. Se revisa el proceso de liberar al paciente del soporte mecánico y de la cánula endotraqueal. El tiempo empleado en el proceso de destete representa de 40 a 50% de la duración total de la ventilación. Se recomienda: 1) Prever tres grupos en función de la dificultad y duración del destete, 2) Retiro lo más pronto posible, 3) Utilizar la prueba de ventilación espontánea para determinar si los pacientes pueden ser extubados con éxito, 4) La prueba inicial debe durar 30 minutos en respiración en tubo en T o con bajos niveles de soporte, 5) La presión de soporte y los modos de ventilación AC deben preferirse cuando han fracasado en la primera prueba, 6) Considerar ventilación no invasiva en pacientes seleccionados para acortar la duración de la intubación, pero no debe usarse de forma rutinaria como herramienta para el fracaso de la extubación.
Abstract: The majority of patients requiring ventilation> 24 hours and improving can be extubated after the first spontaneous ventilation test. The challenge is to improve the disconnection of patients who fail. The withdrawal of mechanical ventilation is an essential element in the care of critically ill patients. The process of releasing the patient from the mechanical support and the endotracheal cannula is reviewed. The time spent in the weaning process represents 40 to 50% of the total duration of ventilation. It is recommended: 1) Predict three groups depending on the difficulty and duration of weaning, 2) Removal as soon as possible, 3) Using the spontaneous ventilation test to determine if patients can be successfully extubated, 4) The initial test Should last 30 minutes in T-tube breathing or with low levels of support, 5) Support pressure and modes of AC ventilation should be preferred when they failed in the first test, 6) Consider non-invasive ventilation in selected patients to shorten The duration of intubation, but should not be routinely used as a tool for the failure of extubation.
Resumo: A maioria dos pacientes que necessitam de ventilação > 24 horas e melhoram podem ser extubados depois do primeiro teste de ventilação espontânea. O desafio é melhorar a desconexão dos pacientes que fracassam. A retirada da ventilação mecânica é um elemento essencial no tratamento dos pacientes em estado crítico. Revisamos o processo de retirada do paciente do suporte mecânico e do tubo endotraqueal. O tempo gasto no processo do desmame representa 40 a 50% da duração total da ventilação. Recomenda-se: 1) Prever três grupos, dependendo da dificuldade e duração do desmame, 2) remoção o mais rápido possível, 3) Utilização do teste de ventilação espontânea para determinar se os pacientes podem ser extubadas com êxito, 4) O teste inicial deve durar 30 minutos com respiração em tubo T ou com baixos níveis de suporte, 5) A pressão de suporte e os modos de ventilação AC deve ser eleitos quando existe uma falha no primeiro teste, 6) Considerar a ventilação não invasiva em pacientes selecionados para diminuir a duração da intubação, mas não deve ser usado rotineiramente como uma ferramenta para falha na extubação.