ABSTRACT
Stable transgenesis is a transformative tool in model organism biology. Although the sea urchin is one of the oldest animal models in cell and developmental biology, studies in this animal have largely relied on transient manipulation of wild animals, without a strategy for stable transgenesis. Here, we build on recent progress to develop a more genetically tractable sea urchin species, Lytechinus pictus, and establish a robust transgene integration method. Three commonly used transposons (Minos, Tol2 and piggyBac) were tested for non-autonomous transposition, using plasmids containing a polyubiquitin promoter upstream of a H2B-mCerulean nuclear marker. Minos was the only transposable element that resulted in significant expression beyond metamorphosis. F0 animals were raised to sexual maturity, and spawned to determine germline integration and transgene inheritance frequency, and to characterize expression patterns of the transgene in F1 progeny. The results demonstrate transgene transmission through the germline, the first example of a germline transgenic sea urchin and, indeed, of any echinoderm. This milestone paves the way for the generation of diverse transgenic resources that will dramatically enhance the utility, reproducibility and efficiency of sea urchin research.
Subject(s)
Animals, Genetically Modified , DNA Transposable Elements , Gene Transfer Techniques , Germ Cells , Lytechinus , Transgenes , Animals , DNA Transposable Elements/genetics , Germ Cells/metabolism , Lytechinus/genetics , Female , Male , Sea Urchins/genetics , Mitochondria Associated MembranesABSTRACT
The identification and characterization of noncanonical functions within the autophagy pathway have unveiled intricate cellular processes, including LC3-associated phagocytosis (LAP) and LC3-associated endocytosis (LANDO). These phenomena play pivotal roles in the conjugation of ATG8 with single-membrane phagosomes and endosomes, shedding light on the dynamic interplay between autophagy and cellular homeostasis. Here, we present detailed protocols for both qualitative and quantitative assessment of LAP, including immunofluorescence, flow cytometry, and Western blotting of isolated LAPosomes. Additionally, the protocol for the evaluation of LANDO through immunofluorescent detection of receptor recycling is outlined. The methodologies presented herein serve as a practical guide for researchers seeking to unravel the intricacies of LAP and LANDO. By providing step-by-step instructions, accompanied by insights into potential challenges and optimization strategies, this chapter aims to empower investigators in the exploration of these noncanonical functions of autophagy proteins.
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T cell exhaustion is linked to persistent antigen exposure and perturbed activation events, correlating with poor disease prognosis. Tumor-mediated T cell exhaustion is well documented; however, how the nutrient-deprived tumor niche affects T cell receptor (TCR) activation is largely unclear. We show that methionine metabolism licenses optimal TCR signaling by regulating the protein arginine methylome, and limiting methionine availability during early TCR signaling promotes subsequent T cell exhaustion. We discovered a novel arginine methylation of a Ca 2+ -activated potassium transporter, KCa3.1, prevention of which results in increased Ca 2+ -mediated NFAT1 activation, NFAT1 promoter occupancy, and T cell exhaustion. Furthermore, methionine supplementation reduces nuclear NFAT1 in tumor-infiltrating T cells and augments their anti-tumor activity. These findings demonstrate metabolic regulation of T cell exhaustion determined during TCR engagement.
ABSTRACT
Pressure ulcers carry a significant risk in clinical practice. This paper proposes a practical and interpretable approach to estimate the risk levels of pressure ulcers using decision tree models. In order to address the common problem of imbalanced learning in nursing classification datasets, various oversampling configurations are analyzed to improve the data quality prior to modeling. The decision trees built are based on three easily identifiable and clinically relevant pressure ulcer risk indicators: mobility, activity, and skin moisture. Additionally, this research introduces a novel tabular visualization method to enhance the usability of the decision trees in clinical practice. Thus, the primary aim of this approach is to provide nursing professionals with valuable insights for assessing the potential risk levels of pressure ulcers, which could support their decision-making and allow, for example, the application of suitable preventive measures tailored to each patient's requirements. The interpretability of the models proposed and their performance, evaluated through stratified cross-validation, make them a helpful tool for nursing care in estimating the pressure ulcer risk level.
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Watermelon (Citrullus lanatus) is a key horticultural crop in Georgia with farmgate value of 142 million USD (2022 Farm Gate Value Report), yet faces challenges from whitefly-transmitted viruses, especially during periods of elevated whitefly populations in the fall. Foliar symptoms on watermelon plants including yellow mottling and chlorosis, wrinkling, bunching, and upward curling, were observed in experimental fields at UGA Tifton and commercial fields in Colquitt County in the fall of 2023. These were similar to those described for watermelon crinkle leaf-associated viruses (WCLaV-1 and WCLaV-2) from Florida (Hendrick et al, 2021) and Texas (Hernandez et al., 2021). The disease incidence reached 100% in both locations. WCLaV-1 was previously identified in Georgia (Adeleke et al., 2022a); however, WCLaV-2 remained undetected in further surveys (Adeleke et al., 2022b). Total nucleic acid was extracted from symptomatic leaf tissues with the MagMAXTM 96 Viral RNA isolation kit (ThermoFisher Scientific, USA), following the manufacturer's guidelines, with the omission of DNAse treatment. The presence of WCLaV-1 was identified through reverse transcription-polymerase chain reaction (RT-PCR) (Hernandez et al., 2021) in 17 out of 24 samples in Tift County, and 12 out of 15 samples from Colquitt County. Within the same set of samples, WCLaV-2 was identified in three samples from Tift County and four samples from Colquitt County by RT-PCR directed at the RdRp gene (Hernandez et al., 2021). WCLaV-1 was not detected in the three samples from Tift County that had WCLaV-2, while mixed infections of WCLaV-2 and WCLaV-1 were observed in the samples form Colquitt County. Two whitefly transmitted viruses, previously reported in Georgia were also identified as mixed infection in these samples (Table S1). The presence of WCLaV-2 was further confirmed by amplifying the movement protein (MP) gene of WCLaV-2 by RT-PCR assays (Hernandez et al., 2021). The amplicons, with expected sizes of 968bp for the RdRp gene and 562bp for the MP gene of WCLaV-2, located on RNA 1 and RNA 2 segments respectively, were directly sequenced from both directions (Genewiz, USA) from a sample collected in Tift County. The resulting data were analyzed via BLASTn search. The MP gene fragment (PP178543) shared 100% identity with isolates from Brazil (LC636074.1), Texas (MW559086.1), and Florida (MZ325858.1). RdRp gene (PP178542) shared >99.7% identity with isolates from Brazil (LC636073.1), Texas (MW559083.1) and Florida (MZ325855.1). WCLaV-1 and WCLaV-2, initially discovered in Asia (Xin et al., 2017), have been assigned to the genus Coguvirus, in the family Phenuiviridae (Walker et al., 2022). Subsequent reports from the USA (Hendrick et al., 2021; Hernandez et al., 2021), Australia (Mulholland et al., 2023), and Brazil (Maeda et al., 2022) indicate the global spread of these viruses. Watermelon is the primary host of WCLaV-2. Despite these findings, biological information, including vector relations, for both viruses and other members of the genus Coguvirus remains elusive. The impact of these viruses on watermelon production and yield in the identified regions remains largely unknown, underscoring the need for further investigations.
ABSTRACT
Background: At the beginning of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, transfusion of coronavirus disease 2019 (COVID-19) convalescent plasma (CCP) emerged as a potential therapeutic strategy to help patients severely afflicted by COVID-19. The efficacy of CCP has been controversial as it depends on many variables pertaining to the plasma donor and the patient with COVID-19, for example, time of convalescence or symptoms onset. This feasibility and descriptive study aimed to assess the safety of multiple doses of CCP in mechanically ventilated, intubated patients with respiratory failure due to COVID-19. Methods: A cohort of 30 patients all experiencing severe respiratory failure and undergoing invasive mechanical ventilation in an intensive care unit, received up to five doses of 300-600 mL of CCP on alternate days (0, 2, 4, 6, and 8) until extubation, futility, or death. Results: Nineteen patients received five doses, seven received four, and four received two or three doses. At 28-day follow-up mark, 57% of patients recovered and were sent home, and the long-term mortality rate was 27%. Ten severe adverse events reported in the study were unrelated to CCP transfusion. Independent of the number of transfused doses, most patients had detectable levels of total and neutralizing antibodies in plasma. Conclusion: This study suggests that transfusion of multiple doses of CCP is safe. This strategy may represent a viable option for future studies, given the potential benefit of CCP transfusions during the early stages of infection in unvaccinated populations and in settings where monoclonal antibodies or antivirals are contraindicated or unavailable.
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OBJECTIVE: The recommendations of the Spanish Ministry of Health on vaccination in risk groups include mesalazine among the treatments with a possible negative effect on its effectiveness. However, this is not the recommendation of most experts. Our objective was to evaluate the effect of mesalazine on the humoral response to the SARS-CoV-2 vaccine in patients with inflammatory bowel disease (IBD). METHODS: VACOVEII is a Spanish, prospective, multicenter study promoted by GETECCU, which evaluates the effectiveness of the SARS-CoV-2 vaccine in patients with IBD. This study includes IBD patients who have recieved the full vaccination schedule and without previous COVID-19 infection. Seroconversion was set at 260BAU/mL (centralized determination) and was assessed 6 months after full vaccination. In this subanalysis of the study, we compare the effectiveness of the vaccine between patients treated with mesalazine and patients without treatment. RESULTS: A total of 124 patients without immunosuppressive therapy were included, of which 32 did not receive any treatment and 92 received only mesalazine. Six months after full vaccination, no significant differences are observed in the mean concentrations of IgG anti-S between both groups. In the multivariate analysis, antibody titers were independently associated with the use of mRNA vaccines and with SARS-CoV-2 infection. CONCLUSION: Mesalazine does not have a negative effect on the response to SARS-CoV-2 vaccines in IBD patients.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal , COVID-19 Vaccines , COVID-19 , Inflammatory Bowel Diseases , Mesalamine , Humans , Mesalamine/therapeutic use , Female , Prospective Studies , Male , COVID-19 Vaccines/immunology , Inflammatory Bowel Diseases/drug therapy , Middle Aged , Adult , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , COVID-19/prevention & control , COVID-19/immunology , Antibodies, Viral/blood , Vaccination , Aged , Seroconversion , Vaccine Efficacy , SARS-CoV-2/immunologyABSTRACT
Fundamentos: Las úlceras por presión (UPP) son las lesiones más frecuentes de las relacionadas con la dependencia y tienenun gran impacto en la calidad de vida de quienes las sufren. Disponer de herramientas en español específicas para evaluar la calidadde vida percibida por los pacientes con UPP se considera un elemento indispensable para la toma de decisiones sobre los cuidadosde salud. El objetivo de este estudio fue traducir al español y adaptar culturalmente el cuestionarioPressure Ulcer Quality of LifeQuestionnaire (PU-QOL) para la medición de la calidad de vida relacionada con la salud en pacientes con úlceras por presión.Métodos: Se utilizó un método de traducción, retrotraducción y pretest a la población diana para obtener una versión adaptadadel instrumento original PU-QOL. El emplazamiento fue la Atención Primaria. Los participantes fueron quince pacientes de AtenciónPrimaria. Mediciones principales: 1) Traducción directa; 2) Síntesis y conciliación de las versiones por un comité de expertos; 3) Tra-ducción inversa; 4) Conciliación de la traducción inversa con la autora del cuestionario original; y 5) Análisis de la comprensibilidadmediante entrevistas cognitivas a una muestra de pacientes.Resultados: Se obtuvo un instrumento de medición de calidad de vida percibida en pacientes con UPP, compuesto por diez es-calas y ochenta y tres ítems. Se mantuvieron las escalas e ítems del cuestionario original. El análisis conceptual y semántico produjoajustes de redacción, clarificación y reformulaciones adaptadas al contexto español.Conclusiones: La disponibilidad de un cuestionario en español de medición de calidad de vida percibida, como es el PU-QOL, delque presentamos esta primera fase de traducción y adaptación transcultural, podrá ser un elemento útil para la toma de decisionessobre los cuidados en salud en pacientes con UPP.(AU)
Background: Pressure ulcers (PU) are the most prevalent of the dependency-related injuries, affecting the quality of life of thepatients who suffer them. However, there are no instruments adapted to the Spanish context to evaluate this quality of life. The useof specific tools in Spanish to evaluate the quality of life perceived by patients with PUs is considered an indispensable element forhealthcare decisions. The aim of this paper was to translate and culturally adapt the Pressure Ulcer Quality of Life Questionnaire (PU-QOL) into Spanish for the measurement of health-related quality of life in patients with pressure ulcers.Methods: A translation, back-translation, and pre-test method was used on the target population to obtain an adapted version of theoriginal PU-QOL instrument. The area was Primary Care. The participants were fifteen Primary Care patients. Main steps: 1) Direct transla-tion; 2) Synthesis and concordance of versions by an expert committee; 3) Back translation; 4) Consistency of the back translation withthe author from the original questionnaire; and 5) Analysis of comprehensibility through cognitive interviews with a sample of patients.Results: An instrument to measure perceived quality of life in patients with PU was obtained, composed of ten scales andeighty-three items. The scales and items of the original questionnaire were maintained. Conceptual and semantic analysis producedadjustments in wording, clarification and reformulations adapted to the Spanish context.Conclusions: We present this first phase of translation and cross-cultural adaptation of the PU-QOL questionnaire in Spanish,which could be a useful tool for decision-making on health care in patients with PUs.(AU)
Subject(s)
Humans , Cultural Diffusion , Quality of Life , Surveys and Questionnaires , Pressure UlcerABSTRACT
No disponible
Subject(s)
Humans , Middle Aged , Aged , Aged, 80 and over , Venous Thromboembolism , Pulmonary Embolism , Venous Thrombosis , Hemorrhage , Retrospective Studies , Vena Cava FiltersABSTRACT
Antecedentes: la hepatitis aguda por virus de la hepatitis E (VHE) en Europa era tradicionalmente una infección adquirida por personas que viajaban a zonas endémicas, fundamentalmente Asia y África. Actualmente, cada vez es mayor el número de casos autóctonos esporádicos diagnosticados en Occidente. Objetivo: analizar los casos diagnosticados de hepatitis aguda por virus hepatitis E (VHE) en nuestro medio, identificando las características clínicas-epidemiológicas. Material y método: se recogieron los casos diagnosticados de hepatitis aguda por VHE (IgM anti-VHE positiva y/o ARN-VHE presente en suero) desde enero de 2008 a diciembre 2014. Se analizaron diversas variables clínicas y epidemiológicas y la evolución posterior. Resultados: se encontraron 23 casos, todos ellos naturales de España. Catorce sujetos (60,87%) presentaban ictericia en el momento del diagnóstico y marcada citolisis (aspartato aminotransferasa [AST] 1.106,91 U/l y alanina aminotransferasa [ALT] 1.407,04 U/l). Veintidós casos fueron considerados autóctonos y uno había realizado un viaje a China tres meses antes. El tiempo medio de resolución fue de 11,2 semanas. En total, diez pacientes (43,5%) mostraban algún marcador de autoinmunidad positivo. Dos sujetos estaban diagnosticados de enfermedad hepática crónica previa y fueron catalogados como 'acute-on-chronic liver failure' (ACLF); uno de ellos finalizó en exitus y el otro, en trasplante hepático. Conclusión: la hepatitis aguda por VHE en nuestro medio es una entidad autóctona, probablemente infradiagnosticada, que se manifiesta con ictericia y citolisis. La presencia de marcadores de autoinmunidad positivos supone un epifenómeno que en ocasiones dificulta su diagnóstico (AU)
Background: In Europe, acute hepatitis caused by the hepatitis E virus (HEV) traditionally was an infection found in people who had travelled to endemic zones, mainly Asia and Africa. However, a growing number of sporadic autochthonous cases are now being diagnosed in the Western world. Objective: To analyze the cases of acute HEV hepatitis diagnosed in our setting, with the identification of the clinical-epidemiological characteristics. Material and methods: We included the cases of acute HEV hepatitis diagnosed (positive anti-HEV IgM and/or HEV RNA present in serum) between January 2008 and December 2014. Different clinical, epidemiological and evolutive parameters were analyzed. Results: A total of 23 patients were identified, all originating from Spain. Fourteen cases (60.87%) presented jaundice and marked cytolysis at the time of diagnosis (aspartate aminotransferase [AST] 1,106.91 U/l and alanine aminotransferase [ALT] 1,407.04 U/l). Twenty-two cases were regarded as autochthonous, and one patient had travelled to China three months before. The mean time to resolution was 11.2 weeks. Some autoimmune markers were positive in 43.5% of the patients. Two subjects were diagnosed with previous chronic liver disease and were classified as 'acute-on-chronic liver failure' (ACLF), one died and the other underwent liver transplantation. Conclusion: Acute HEV hepatitis in our setting is an autochthonous condition that is probably underdiagnosed, manifesting with jaundice and cytolysis. Autoimmune marker positivity is an epiphenomenon, which in some cases complicates the diagnosis (AU)
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Hepatitis E/diagnosis , Hepatitis E/epidemiology , Jaundice/complications , Autoimmunity/physiology , Hepatitis A/diagnosis , Hepatitis, Autoimmune/diagnosis , Retrospective Studies , Immunoglobulin M/analysis , Immunoglobulin G/analysis , Enzyme-Linked Immunosorbent Assay/methods , Liver Failure/complications , Diagnosis, DifferentialABSTRACT
El presente trabajo se realizó en pacientes que estuvieron hospitalizados en el Servicio de Neumología y Servicio de Medicina Interna o que acudieron a consultorio externo de neumología del Hospital Regional Honorio Delgado de Arequipa - Perú entre marzo - mayo de 1998. El propósito de nuestro estudio fue evaluar en los pacientes recién diagnosticados de tuberculosis pulmonar severa la alteración de la función ventilatoria en las vías aéreas centrales y periféricas; y la reversibilidad que presenta a la terapia broncodilatadora con sulfato de salbutamol, bromuro de ipratropio y la combinación fija de sulfato de salbutamol más bromuro de ipratropio. Se tomó una problación total de 60 pacientes, de 20 a 40 años de edad, de ambos sexos todos ellos cumplían con los criterios de inclusión. Se formó tres grupos pacientes, los cuales se seleccionaron por orden de llegada, a quienes se les realizó una espirometría (control o basal). Al primer grupo, se les aplicó dos inhalaciones de salbutamol (100 mcg por inhalación), al segundo grupo se le aplicó dos inhalaciones de bromuro de ipratropio (20 mcg por inhalación) y al tercer grupo se aplicó dos inhalaciones de la combinación fija de sulfato de salbutamol más bromuro de ipratropio (120 mcg de sulfato de salbutamol y 20 mcg de bromuro de ipratropio por inhalación), luego de 15 minutos se realizó la segunda espirometría, en cada uno de los casos. El tipo de estudio fue transversal y comparativo. Se empleó pruebas de estadística descriptiva (media aritmética, desviación estándar) y pruebas estadísticas para comprobación de la hipótesis ("t" de student) para establecer la significancia con nivel de P menor 0,05. En nuestros grupos de estudio se observó: los pacientes con tuberculosis pulmonar severa presentan; insuficiencia ventilatoria mixta. El salbutamol y la combinación fija de sulfato de salbutamol más bromuro de ipratropio, actúan significativamente en la reversibilidad de las vías aéreas periféricas. El bromuro de ipratropio presenta respuesta broncodilatadora significativa en las vías centrales y periféricas, predominando en la primera.
Subject(s)
Humans , Male , Female , Adult , Albuterol , Bromides , Ipratropium , Spirometry , Sulfates , Tuberculosis, Pulmonary/therapy , Cross-Sectional StudiesABSTRACT
El presente trabajo se realizó en el área ambulatoria del Servicio de Neumología del Hospital Regional Honorio Delgado de Arequipa-Perú desde el 1 de Enero hasta el 31 de Agosto de 1997, considerándose a todos los pacientes con asma bronquial intercrítico que reunían los criterios de inclusión para ser considerados en el presente estudio. El objetivo de nuestra investigación se centró en precisar sí el efecto broncodilatador del salbutamol era más, o menos intenso, en los pacientes con asma intercrítico en relación al uso de la asociación salbutamol bromuro de ipratropio y además precisar en cúal de las vías aéreas se objetivaba una mejor acción con uno u otro recurso terapeútico. En el presente trabajo se determinó a través del estudio de la capacidad vital forzada y volumen espiratorio forzado al segundo, que el test de broncodilatación, utilizando un anticolinérgico como bromuro de ipratropio más salbutamol, es más sensible que el test del salbutamol sólo para el diagnóstico de asma bronquial intercrítico. El compromiso de las vías aéreas centrales pueden ser mejor determinadas por la cuantificación del pico flujo máximo y de manera más específica con resultados confiables estudiando al flujo espiratorio forzado 25 por ciento, éste último parámetro sólo se torna fiable cuando se realiza la doble broncodilatación con el bromuro de ipratropio y salbutamol simultáneamente. Las vías aéreas periféricas son mejor estudiadas y su compromiso mejor cuantificado cuando determinamos el flujo espiratorio forzado 25-75 y la posibilidad de error se toma mucho menor cuando se utiliza el test de la doble broncodilatación.
Subject(s)
Albuterol , Asthma , Ipratropium , SpirometryABSTRACT
El presente trabajo se realizó en pacientes que fueron atendidos en el consultorio externo de Neumología del Hospital Regional "Honorio Delgado" de Arequipa - Perú, del 1§ de Enero al 31 de Diciembre de 1996 a quienes se les hizo el diagnóstico de asma bronquial tanto clínica como laboratorialmente, utilizando para ello ademas del cuadro clínico compatible, el estudio espirométrico con curva flujo volumen y test de salbutamol que indicaba reversibilidad del cuadro obstructivo bronquial; fueron excluidos del presente estudio los pacientes que no tuvieron estudio laboratorial positivo o que al momento de realizar la evaluación espirométrica se onjetivó una insuficiencia ventilatoria restrictiva que hacía pensar en otro tipo de patología. El propósito del presente trabajo fue determinar en qué magnitud se afectaban tanto las vías aéreas centrales (gruesas), como las periféricas (delgadas) o ambas a la vez, en pacientes asmáticos en relación al sexo y a la contaminación ambiental y a que viven a mediana altura. Se tomó una población total de 94 pacientes, siendo considerados en el trabajo 68 consultantes que cumplían con los criterios de inclusión. Se observó que en nuestro grupo de estudio se comprometen por igual, tanto las vías aéreas centrales como las periféricas en hombres y mujeres, por lo que pensamos que un tratamiento coherente para un paciente asmático es aquel que incluye además de la terapia inhalatoria corticoidea, un beta 2 agonista que tiene mejor en las vías aéreas centrales.
Subject(s)
Airway Obstruction , Asthma/diagnosis , Asthma/pathology , SpirometryABSTRACT
En esta segunda edición, el autor sigue planteando al presente, problemas no resueltos y alternativas de sorprendente vigencia, ofreciéndonos una visión del Perú, cincuenta y cinco años después. Propone un descentralismo productivo que resuelva problemas del centralismo, la pobreza y la ausencia de planificación; así también plantea la tesis precursora de combinar regiones transversales y horizontales considerando el mar y los ríos no como fronteras sino como elementos claves de dicha articulación