RESUMO
BACKGROUND: Developing effective vaccines against SARS-CoV-2 that consider manufacturing limitations, equitable access, and acceptance is necessary for developing platforms to produce antigens that can be efficiently presented for generating neutralizing antibodies and as a model for new vaccines. RESULTS: This work presents the development of an applicable technology through the oral administration of the SARS-CoV-2 RBD antigen fused with a peptide to improve its antigenic presentation. We focused on the development and production of the recombinant receptor binding domain (RBD) produced in E. coli modified with the addition of amino acids extension designed to improve antigen presentation. The production was carried out in shake flask and bioreactor cultures, obtaining around 200 mg/L of the antigen. The peptide-fused RBD and peptide-free RBD proteins were characterized and compared using SDS-PAGE gel, high-performance chromatography, and circular dichroism. The peptide-fused RBD was formulated in an oil-in-water emulsion for oral mice immunization. The peptide-fused RBD, compared to RBD, induced robust IgG production in mice, capable of recognizing the recombinant RBD in Enzyme-linked immunosorbent assays. In addition, the peptide-fused RBD generated neutralizing antibodies in the sera of the dosed mice. The formulation showed no reactive episodes and no changes in temperature or vomiting. CONCLUSIONS: Our study demonstrated the effectiveness of the designed peptide added to the RBD to improve antigen immunostimulation by oral administration.
Assuntos
COVID-19 , SARS-CoV-2 , Animais , Humanos , Camundongos , Adjuvantes Imunológicos , Vacinas contra COVID-19 , Escherichia coli , Administração Oral , Antígenos Virais , Anticorpos Neutralizantes , Peptídeos , Anticorpos AntiviraisRESUMO
Resumo A gestação tem recebido pouca atenção quanto às modificações normais percebidas nos domínios físicos e psicológicos do estado da saúde da mulher e de sua qualidade de vida, embora seja um evento comum na vida reprodutiva feminina. O objetivo deste estudo foi avaliar a qualidade de vida e as estratégias de enfrentamento das gestantes de alto risco e risco habitual, do segundo e do terceiro trimestres de gestação, do município de Dourados, Mato Grosso do Sul, Brasil. Participaram 79 gestantes, das quais 32 eram de alto risco e 47, de risco habitual, com idade média de 24,6 anos; aplicou-se um questionário de qualidade de vida e outro de enfrentamento. As gestantes de alto risco apresentam maior qualidade de vida nos domínios saúde/funcionamento e família; desse modo, em todos os fatores significativos de enfrentamento. Observa-se a necessidade de acompanhamento dessas participantes em programas especiais inseridos no pré-natal, a fim de contribuir para o enfrentamento da gestação.
Resumen La gestación ha recibido poca atención en cuanto a las modificaciones percebidas en los dominios físico y psicológico del estado de la salud de la mujer y su calidad de vida, aun siendo un evento común en la vida reproductiva femenina. El propósito del estudio fue evaluar la calidad de vida y las estrategias de afrontamiento de gestantes de alto y habitual riesgo durante el segundo y tercer trimestres de gestación en el municipio de Dorados (Mato Grosso do Sul, Brasil). Participaron 79 gestantes, siendo 32 de alto riesgo y 47 de riesgo habitual, con promedio de edad de 24,6 años; a quienes se les aplicó un cuestionario de calidad de vida y otro de enfrentamiento. Las embarazadas de alto riesgo presentan mayor calidad de vida en los dominios de Salud/ Funcionamiento y Familia; asimismo, en todos los factores significativos de enfrentamiento. Se observa la necesidad de acompañamiento de las participantes en programas especiales que son incluidos en el prenatal con el fin de contribuir al enfrentamiento de la gestación.
Abstract Pregnancy has received little attention in terms of the perceived changes in the physical and psychological domain of women's health status and quality of life. The purpose of this study was to evaluate the quality of life and coping strategies of pregnant women during the second and third trimesters of pregnancy, in the municipality of Dourados (Mato Grosso do Sul). A total of 79 pregnant women participated in the study, 32 of whom were high-risk and 47 average risk, with an average age of 24.6 years; to whom a quality of life questionnaire and a coping questionnaire were applied. High-risk pregnant women have higher quality of life in the Health/Functioning and Family domains; likewise, in all the significant coping factors. It is observed the need to accompany participants in special programs that are included in the prenatal period in order to contribute to coping with pregnancy.
RESUMO
Chronic consumption of ß-sitosterol-ß-D-glucoside (BSSG), a neurotoxin contained in cycad seeds, leads to Parkinson's disease in humans and rodents. Here, we explored whether a single intranigral administration of BSSG triggers neuroinflammation and neurotoxic A1 reactive astrocytes besides dopaminergic neurodegeneration. We injected 6 µg BSSG/1 µL DMSO or vehicle into the left substantia nigra and immunostained with antibodies against tyrosine hydroxylase (TH) together with markers of microglia (OX42), astrocytes (GFAP, S100ß, C3), and leukocytes (CD45). We also measured nitric oxide (NO), lipid peroxidation (LPX), and proinflammatory cytokines (TNF-α, IL-1ß, IL-6). The Evans blue assay was used to explore the blood-brain barrier (BBB) permeability. We found that BSSG activates NO production on days 15 and 30 and LPX on day 120. Throughout the study, high levels of TNF-α were present in BSSG-treated animals, whereas IL-1ß was induced until day 60 and IL-6 until day 30. Immunoreactivity of activated microglia (899.0 ± 80.20%) and reactive astrocytes (651.50 ± 11.28%) progressively increased until day 30 and then decreased to remain 251.2 ± 48.8% (microglia) and 91.02 ± 39.8 (astrocytes) higher over controls on day 120. C3(+) cells were also GFAP and S100ß immunoreactive, showing they were neurotoxic A1 reactive astrocytes. BBB remained permeable until day 15 when immune cell infiltration was maximum. TH immunoreactivity progressively declined, reaching 83.6 ± 1.8% reduction on day 120. Our data show that BSSG acute administration causes chronic neuroinflammation mediated by activated microglia, neurotoxic A1 reactive astrocytes, and infiltrated immune cells. The severe neuroinflammation might trigger Parkinson's disease in BSSG intoxication.
Assuntos
Astrócitos/efeitos dos fármacos , Astrócitos/imunologia , Inflamação/etiologia , Neurotoxinas/imunologia , Sitosteroides/administração & dosagem , Substância Negra/efeitos dos fármacos , Substância Negra/metabolismo , Animais , Astrócitos/metabolismo , Biomarcadores , Doença Crônica , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Leucócitos/imunologia , Leucócitos/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Microglia/imunologia , Microglia/metabolismo , Neurotoxinas/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Doença de Parkinson/etiologia , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Ratos , Substância Negra/patologiaRESUMO
Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/ß pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/ß pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy.
Assuntos
Portadores de Fármacos/síntese química , Técnicas de Transferência de Genes , Vetores Genéticos/administração & dosagem , Neurotensina/administração & dosagem , Fragmentos de Peptídeos/síntese química , Transporte Ativo do Núcleo Celular , Animais , Linhagem Celular , Núcleo Celular/metabolismo , Neurônios Dopaminérgicos/citologia , Neurônios Dopaminérgicos/metabolismo , Terapia Genética/métodos , Vetores Genéticos/genética , Masculino , Camundongos , Modelos Animais , Nanopartículas/química , Neurotensina/genética , Neurotensina/farmacocinética , Técnicas de Patch-Clamp , Plasmídeos/genética , Ratos , Receptores de Neurotensina/metabolismo , Análise de Célula Única , Técnicas EstereotáxicasRESUMO
The spreading and accumulation of α-synuclein and dopaminergic neurodegeneration, two hallmarks of Parkinson's disease (PD), have been faithfully reproduced in rodent brains by chronic, oral administration of ß-sitosterol ß-D-glucoside (BSSG). We investigated whether a single injection of BSSG (6 µg BSSG/µL DMSO) in the left substantia nigra of Wistar rats causes the same effects. Mock DMSO injections and untreated rats formed control groups. We performed immunostainings against the pathological α-synuclein, the dopaminergic marker tyrosine hydroxylase (TH), the neuroskeleton marker ß-III tubulin, the neurotensin receptor type 1 (NTSR1) as non-dopaminergic phenotype marker and Fluro-Jade C (F-J C) label for neurodegeneration. Using ß-galactosidase (ß-Gal) assay and active caspase-3 immunostaining, we assessed cell death mechanisms. Golgi-Cox staining was used to measure the density and types of dendritic spines of striatal medium spiny neurons. Motor and non-motor alterations were also evaluated. The study period comprised 15 to 120 days after the lesion. In the injured substantia nigra, BSSG caused a progressive α-synuclein aggregation and dopaminergic neurodegeneration caused by senescence and apoptosis. The α-synuclein immunoreactivity was also present within microglia cells. Decreased density of dopaminergic fibers and dendritic spines also occurred in the striatum. Remarkably, all the histopathological changes also appeared on the contralateral nigrostriatal system, and α-synuclein aggregates were present in other brain regions. Motor and non-motor behavioral alterations were progressive. Our data show that the stereotaxic BSSG administration reproduces PD α-synucleinopathy phenotype in the rat. This approach will aid in identifying the spread mechanism of α-synuclein pathology and validate anti-synucleinopathy therapies.
Assuntos
Modelos Animais de Doenças , Degeneração Neural/patologia , Doença de Parkinson , Sitosteroides/administração & dosagem , alfa-Sinucleína/metabolismo , Animais , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/patologia , Injeções Intraventriculares/métodos , Degeneração Neural/induzido quimicamente , Ratos , Ratos Wistar , Sitosteroides/toxicidade , Substância Negra/efeitos dos fármacos , Substância Negra/patologiaRESUMO
Las fracturas del hueso ganchoso son muy poco frecuentes, constituyendo alrededor del 2% de todas las fracturas del carpo. Debido a su baja frecuencia y a lo inespecífico de las manifestaciones clínicas, su diagnóstico suele ser difícil. Es importante considerar su presencia ante un traumatismo directo sobre la mano. Debe estudiarse inicialmente con radiografía de mano. Si es necesario confirmar el diagnóstico o valorar las lesiones asociadas, se sugiere la tomografía axial computarizada. La decisión terapéutica depende del grado de conminución de la fractura, de su estabilidad y de la presencia de compromiso articular. Se presenta el caso de un varón de 19 años que sufrió fractura aislada del cuerpo del hueso ganchoso.
Fractures of the hamate bone are very infrequent, representing approximately 2% of the carpal fractures. Due to its infrequency and to the non-specificity of its clinical manifestations, they are usually difficult to diagnose. Is important to consider its presence when a direct trauma over hand occurs. Study should start with X ray of the hand. If necessary to confirm the diagnosis or assess associated lesions, computed tomography is suggested. The therapeutic management depends on the degree of comminution of the fracture, its stability and any joint involvement. We present a case of a 19 year old male who suffered an isolated fracture of the body of the hamate bone.
RESUMO
Objetivo: Informar los resultados de un estudio piloto que probó la aplicabilidad, eficacia y aceptabilidad de un programa diseñado para prevenir la depresión en estudiantes secundarios. Metodología: El estudio se llevó a cabo en la comuna de San Bernardo. Se usó una metodología mixta: cualitativa y cuantitativa. Se llevó a cabo un ensayo clínico controlado con dos ramas y grupos focales. La muestra estuvo constituida por tres colegios municipales, cuatro clases de 1º Medio en el grupo activo y tres en el grupo control. La intervención de la rama activa consistió en un programa de 11 sesiones, liderado por dos profesionales jóvenes. Resultados: La muestra estuvo constituida por 277 escolares de 1º Medio, 163 en el grupo activo y 114 en el grupo control. La edad promedio de la muestra fue de 14,5 años (DE=0,6). En el diagnóstico basal, el grupo activo no se diferenció significativamente del grupo control en relación a la edad ni al nivel de sintomatología depresiva. En el grupo activo, el puntaje del BDI-II inicialmente fue de 10,7 (IC 95%=9,0 a 12,4), aumentó al término de la intervención a 11,5 (IC 95%=9,7 a 13,4) y a los 6 meses fue de 9,2 (IC 95 por ciento =7,5 a 10,9). En el grupo control, el promedio en el BDI-II fue de 9,2 (IC 95 por ciento =7,6-10,8); 8,0 (IC 95 por ciento =6,3 a 9,6) y 8,6 (IC 95 por ciento =6,1 a 11,1), respectivamente. No hubo diferencias estadísticamente significativas en los resultados entre ambos grupos en la sintomatología depresiva. Sin embargo, la intervención actuó como un factor protector para depresión (OR=0,39; IC 95%=0,19 a 0,79). En la evaluación cualitativa, los participantes calificaron positivamente el taller. Conclusiones: El estudio piloto demostró que es posible aplicar programas de este tipo en los colegios con buena aceptabilidad de los adolescentes. La resultados hasta el seguimiento a 6 meses sugieren que la intervención podría reducir la aparición de nuevos casos de depresión.
Objective: To report the results of a pilot study that tested the applicability, efficacy and acceptability of a program designed to prevent depression among secondary school students. Methodology: The study was conducted in the borough of San Bernardo. A mixed methodology was used: qualitative and quantitative. A randomized controlled trial was carried out with two arms and focus groups. The sample included three state funded schools, four 9th grade classes and three in the control group. The intervention in the active arm was an 11-session program led by two young professionals. Results: The sample constituted 277 students, 163 in the active group and 114 in the control group. The mean age of the sample was 14.5 years (SD=0.6). In the baseline diagnosis, the active group did not differ significantly from the control group regarding age and level of depressive symptoms. In the active group, the BDI-II score was initially 10.7 (95 percent CI=9.0 to 12.4) which increased at the end of the intervention to 11.5 (95 percent CI=9.7 to 13.4) and at the 6 months assessment was 9.2 (95 percent CI=7.5 to 10.9). In the control group, the BDI-II mean score was 9.2 (95 percent CI=7.6 to 10.8); 8.0 (95 percent CI=6.3 to 9.6) and 8.6 (95 percent CI=6.1 to 11.1), respectively. There were no statistically significant differences in the depressive symptoms measured between the control and active groups. However, the intervention acted as a protective factor for depression (OR=0.39; CI 95 percent =0.19 to 0.79). In the qualitative evaluation, participants appraised the program positively. They expressed greater interest in the sessions related to problem-solving strategies. Conclusions: This pilot study demonstrated that it is possible to implement programs like this in schools with good acceptance. The results reached at the time of the 6-month follow-up appear to suggest that the intervention could reduce new cases of depression.
Assuntos
Humanos , Masculino , Adolescente , Feminino , Depressão/prevenção & controle , EstudantesRESUMO
An ethanol extract prepared from the gametophyte Chilean native moss Sphagnum magellanicum was dried out, weighed and dissolved in distilled water. This extract was then assayed for its antibacterial activity against the G(-) bacteria Azotobacter vinelandii, Erwinia carotovora subsp. carotovora, Enterobacter aerogenes, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Vibrio cholerae, and the G(+) bacteria Staphylococcus aureus subsp. aureus, and Streptococcus type beta. The growth of the cultures of E. carotovora subsp. carotovora, and V. cholerae was inhibited at a concentration of 581 microg/ml of extract, while the cultures of E. coli, S. typhi and Streptococcus type beta were inhibited at a concentration of 1.16 microg/mL of extract. The concentration of phenolic compounds was 4.294 mg/mL; the presence of vanillic, chlorogenic, syringic, caffeic, gallic, 3-4 hydrozybenzoic, p-coumaric and salicylic acids was identified using RP- High Pressure Liquid Chromatography.
Assuntos
Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Extratos Vegetais/farmacologia , Sphagnopsida/química , Antibacterianos/isolamento & purificação , Chile , Cromatografia Líquida de Alta Pressão , Bactérias Gram-Negativas/classificação , Bactérias Gram-Positivas/classificação , Testes de Sensibilidade MicrobianaRESUMO
El locus de control es un rasgo de personalidad vinculado con la atribución que hacen los individuos acerca de sus éxitos y fracasos. Los estudios en diferentes naciones y sociedades han demostrado que existen características generales y particulares respecto a la percepción de control y donde la cultura juega un papel importante. Se partió del objetivo de conocer la composición factorial del locus de control en dos poblaciones, Hermosillo, Sonora (México) (n=600) y João Pessoa, Paraíba (Brasil) (n=600); y establecer las diferencias en cuanto a dicho constructo en ambas poblaciones con respecto a las variables atributivas: edad, escolaridad, ocupación y ciudad. Los resultados revelan tres dimensiones comunes a las dos poblaciones: locus de control interno, locus de control afiliativo y locus de control externo. Los datos en general hacen ver que las muestras poblacionales de las ciudades hacen juicios de control en relación a los esfuerzos y el trabajo personal, en la afiliación a colectivos y grupos de interés como los fundamentos de explicación para éxitos y fracasos.
The locus of control is a personality trait linked to the attribution people make concerning their success and failures. Research in nations and societies has demonstrated that there exist general and particular characteristics about control perception, where culture plays an important role. The initial objective was to know the factorial composition about locus of control in two groups located in Hermosillo, Sonora México (n=600) and João Pessoa, Paraíba. Brazil (n=600); then, to establish the differences of the construct between the groups concerning the attribute variables: age, education level, occupation and city. The results show three dimensions in the two groups: internal, affiliation and external locus of control. The general data suggest that the population samples of the cities make control judgements related with efforts and personal work, affiliation to collectives and groups of interest as the basis for explaining their success and failures.
RESUMO
Objetivo: validar en población mexicana la prueba de Allen realizada con oximetría de pulso en pacientes anestesiados. Material y método. Se incluyeron 101 pacientes, de 15 a 73 años de edad. A cada uno se le realizó la prueba de Allen convencional y en forma ciega a los resultados de ésta, detección de la onda de pulso mediante oximetría. Resultados; no se encontró diferencia estadísticamente significativa entre los dos métodos. El tiempo máximo de llenado capilar fue de 2.6 segundos y la correlación entre ambas fue de 0.70. Conclusión: La simplicidad de la técnica descrita y la amplia disponibilidad de un oxímetro en sala de operaciones, hacen de esta prueba una alternativa útil y válida para la evaluación de la circulación de la mano en pacientes anestesiados o con pérdida del estado de conciencia
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Perfusão , Oximetria , Oximetria/instrumentação , Artéria Radial , Mãos/irrigação sanguínea , Cateterismo Periférico , Circulação ColateralRESUMO
Se estudiaron 30 pacientes con enfermedad de Hansen multibacilar (BL o LL) que terminaron el tratamiento controlado según el esquema de la OMS. Se valoraron las baciloscopias de cada paciente en sus índices bacteriológico, morfológico y tintorial durante el inicio, primer año y segundo a lo del tratamiento. Al final del tratamiento 40 por ciento de los pacientes (12) negativizó su índice bacteriológico, el resto de ellos permaneció positivo, por lo que sugerimos que los pacientes con Hansen multibacilar prolonguen por más tiempo el esquema de tratamiento que ha implantado la OMS, para evitar futuras recaídas
Assuntos
Protocolos Clínicos , Tratamento Farmacológico/normas , Tratamento Farmacológico/tendências , Hanseníase/imunologia , Hanseníase/terapia , Organização Mundial da Saúde/organização & administração , Prognóstico , Técnicas BacteriológicasRESUMO
Se informa de una complicación postanestésica no referida en la literatura ocurrida en una paciente de 32 años de edad con Insuficiencia Renal Crónica (IRC), con pruebas de coagulación normales, a quien se les efectuó una anestesia regional con bloqueo epidural para colocar un catéter de Tenchkoff para diálisis peritoneal. En el postoperatorio refiere debilidad y parestesias en las extremidades inferiores y se integra un Síndrome de Neurona Motora Baja, se confirma el diagnóstico de hematoma epidural por imagen de resonancia magnética nuclear a nivel de T12- L1-2 con compresión del cono medular. Se evacúa el hematoma epidural por laminectomía y se descomprime el saco dural. Se observa discreta mejoría en sensibilidad y desaparecen las parestesias en las extremidades inferiores. Recupera movilidad en la extremidad inferior derecha en un 30 por ciento, no así en la izquierda. Se egresa 19 días después de su primera intervención quirúrgica, para continuar con su programa de diálisis peritoneal y rehabilitación física. Se discute el caso y se analizan los trastornos de la coagulación en los pacientes con insuficiencia renal crónica y los peligros potenciales que representa la anestesia regional en este grupo de pacientes. Se destaca la importancia de la resonancia magnética para el apoyo diagnóstico e intervención terapéutica inmediata
Assuntos
Humanos , Feminino , Adulto , Transtornos da Coagulação Sanguínea , Hematoma Epidural Craniano/etiologia , Anestesia por Condução/efeitos adversos , Compressão Nervosa , Insuficiência Renal Crônica/complicações , Espectroscopia de Ressonância Magnética/uso terapêuticoRESUMO
La aplicación clínica de un novedoso método de cobertura, diseñado basándose en la información de la anatomía microvascular de la pierna es presentado, luego de haber sido utilizado en tres pacientes de nuestros servicios clínicos. Junto a la presentación de dichos casos clínicos, la historia, la anatomía macro y microvascular y las relaciones de los diversos componentes que soportan el diseño de este colgajo espresentado, con el único objetivo de mostrar otra alternativa, para la cobertura de este difícil segmentode la anatomía