RESUMEN
Protein-based vaccines are playing an increasingly important role in the COVID-19 pandemic. As late-stage clinical data are finalized and released, the number of protein-based vaccines expected to enter the market will increase significantly. Most protein-based COVID-19 vaccines are based on the SARS-CoV-2 spike protein (S-protein), which plays a major role in viral attachment to human cells and infection. As a result, in order to develop and manufacture quality vaccines consistently, it is imperative to have access to selective and efficient methods for the bioanalytical assessment of S-protein. In this study, samples of recombinant S-protein (hexS-protein) and commercial S-protein were used to develop a selective reversed-phase HPLC (RP-HPLC) method that enabled elution of the intact S-protein monomer as a single peak on a wide pore, C8-bonded chromatographic column. The S-protein subunits, S1 and S2 subunits, were clearly separated from intact S-protein and identified. The results of this study set the foundation for reversed-phase HPLC method development and analysis for selective and efficient separation of S-protein monomer from its subunits.
Asunto(s)
COVID-19 , Glicoproteína de la Espiga del Coronavirus , Vacunas contra la COVID-19 , Cromatografía Líquida de Alta Presión/métodos , Humanos , Pandemias , SARS-CoV-2RESUMEN
Disruptive behaviors can be of comparable or greater concern to parents than the core symptoms of Autism Spectrum Disorder (ASD). Provision of effective interventions to address these behaviors within the first year of initial diagnosis holds great potential for improving the child's, parents', and family's functioning. We piloted a four-session, manualized, positive parenting program on 21 parents of newly diagnosed children ages 2 through 12 years using a mixed methods design. Seventy-five percent of parents completed four sessions, with 100% reporting high levels of service satisfaction. Preliminary results indicated clinically and statistically significant reductions in child maladaptive behaviors, as well as improvements in parental and family functioning. Practitioners and parents identified several potential implementation adaptations, including additional sessions to focus on ASD education and real-time parent-child interactions. Taken as a whole, these data suggest that a brief positive parenting intervention may be a feasible way to improve child, parent, and family functioning during the first year of ASD diagnosis. Findings point to the need for additional research to determine treatment efficacy and to assist with the identification of moderators and mediators of effects.
Asunto(s)
Trastorno del Espectro Autista/terapia , Educación no Profesional/métodos , Relaciones Padres-Hijo , Responsabilidad Parental/psicología , Psicoterapia Breve/métodos , Adulto , Trastorno del Espectro Autista/psicología , Niño , Preescolar , Femenino , Humanos , Masculino , Proyectos Piloto , Evaluación de Programas y Proyectos de SaludRESUMEN
OBJECTIVE: to describe parenting self-efficacy and family empowerment among expectant mothers with substance use disorders. DESIGN: the study employed a quantitative cross-sectional design. PARTICIPANTS: participants were 71 pregnant women with substance use disorders entering a gender-specific, substance use disorder, residential treatment facility. MEASUREMENTS: measurement tools included: the demographic data form, the Parenting Sense of Competence Scale, the Addiction Severity Index, and the Family Empowerment Scale. FINDINGS: women in their third trimester reported statistically significant higher levels of both parenting self-efficacy and family empowerment than women in their first trimester. KEY CONCLUSIONS: interventions that promote parenting self-efficacy and family empowerment need to target women in their first trimester. IMPLICATIONS FOR PRACTICE: midwives with specialized training in substance abuse disorders are in a unique position to bolster expectant mothers' parenting capabilities during the addiction recovery process.
Asunto(s)
Partería , Responsabilidad Parental , Complicaciones del Embarazo/psicología , Apoyo Social , Trastornos Relacionados con Sustancias/psicología , Adulto , Estudios Transversales , Femenino , Humanos , Servicios de Salud Materno-Infantil , Missouri , Embarazo , Escalas de Valoración Psiquiátrica , Tratamiento Domiciliario , Adulto JovenRESUMEN
Oxidative stress is associated with many physiological and pathological processes, as well as xenobiotic metabolism, leading to the oxidation of biomacromolecules, including DNA. Therefore, efficient detection of DNA oxidation is important for a variety of research disciplines, including medicine and toxicology. A common biomarker of oxidatively damaged DNA is 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo; often erroneously referred to as 8-hydroxy-2'-deoxyguanosine (8-OH-dGuo or 8-oxo-dG)). Several protocols for 8-oxo-dGuo measurement by high pressure liquid chromatography with electrochemical detection (HPLC-ED) have been described. However, these were mainly applied to purified DNA treated with pro-oxidants. In addition, due to methodological differences between laboratories, mainly due to differences in analytical equipment, the adoption of published methods for detection of 8-oxo-dGuo by HPLC-ED requires careful optimization by each laboratory. A comprehensive protocol, describing such an optimization process, is lacking. Here, a detailed protocol is described for the detection of 8-oxo-dGuo by HPLC-ED, in DNA from cultured cells or animal tissues. It illustrates how DNA sample preparation can be easily and rapidly optimized to minimize undesirable DNA oxidation that can occur during sample preparation. This protocol shows how to detect 8-oxo-dGuo in cultured human alveolar adenocarcinoma cells (i.e., A549 cells) treated with the oxidizing agent KBrO3, and from the spleen of mice exposed to the polycyclic aromatic hydrocarbon dibenzo(def,p)chrysene (DBC, formerly known as dibenzo(a,l)pyrene, DalP). Overall, this work illustrates how an HPLC-ED methodology can be readily optimized for the detection of 8-oxo-dGuo in biological samples.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , ADN/química , Desoxiguanosina/análogos & derivados , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Biomarcadores/análisis , Biomarcadores/metabolismo , Línea Celular , Células Cultivadas , ADN/metabolismo , Daño del ADN , Desoxiguanosina/análisis , Desoxiguanosina/metabolismo , Humanos , Masculino , Ratones , Oxidantes/química , Oxidantes/farmacología , Oxidación-Reducción , Bazo/química , Bazo/efectos de los fármacosRESUMEN
A flexible synthesis of dibenzo[e,g]isoindol-1-ones has been developed. Dibenzo[e,g]isoindol-1-ones represent simplified benzenoid analogues of biological indolo[2,3-a]pyrrolo[3,4-c]carbazol-5-ones (indolocarbazoles), compounds that have demonstrated a wide range of biological activity. The synthesis of the title compounds involved tetramic acid sulfonates. Different aryl groups were introduced at C4 of the heterocyclic ring via Suzuki-Miyaura cross-coupling reactions. Finally, mild Scholl-type oxidative cyclizations mediated by phenyliodine(III) bis(trifluoroacetate) (PIFA) converted some of the latter compounds into the corresponding dibenzo[e,g]isoindol-1-ones. A systematic study of the oxidative cyclization revealed the following reactivity trend: 3,4-dimethoxyphenyl â« 3-methoxyphenyl > 3,4,5-trimethoxyphenyl > 4-methoxyphenyl ≈ phenyl. Overall, the oxidative cyclization required at least two methoxy groups distributed in the aromatic rings, at least one of which had to be located para to the site of the cyclization.