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Ivacaftor is the first clinically approved monotherapy potentiator to treat CFTR channel dysfunction in people with cystic fibrosis. Ivacaftor (Iva) is a critical component for all current modulator therapies, including highly effective modulator therapies. Clinical studies show that CF patients on ivacaftor-containing therapies present various clinical responses, off-target effects, and adverse reactions, which could be related to metabolites of the compound. In this study, we reported the concentrations of Iva and two of its major metabolites (M1-Iva and M6-Iva) in capillary plasma and estimated M1-Iva and M6-Iva metabolic activity via the metabolite parent ratio in capillary plasma over 12 h. We also used the ratio of capillary plasma versus human nasal epithelial cell concentrations to evaluate entry into epithelial cells in vivo. M6-Iva was rarely detected by LC-MS/MS in epithelial cells from participants taking ivacaftor, although it was detected in plasma. To further explore this discrepancy, we performed in vitro studies, which showed that M1-Iva, but not M6-Iva, readily crossed 16HBE cell membranes. Our studies also suggest that metabolism of these compounds is unlikely to occur in airway epithelia despite evidence of expression of metabolism enzymes. Overall, our data provide evidence that there are differences between capillary and cellular concentrations of these compounds that may inform future studies of clinical response and off-target effects.
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Elexacaftor, tezacaftor, ivacaftor (ETI) is a CFTR modulator combination approved for use in â¼90 % of people with cystic fibrosis (pwCF) over 2 years old. While most pwCF tolerate this therapy well, some are intolerant to standard dosing, and others show little response. Clinical providers may adjust ETI dosing to combat these issues, but these adjustments are not well guided by pharmacokinetic evidence. Our post-approval study aimed to describe pharmacokinetic variability of ETI plasma concentrations in 15 participants who were administered a standard or reduced dose. ETI were quantified by LC-MS/MS in plasma samples taken prior to the morning dose. Results showed non-significant differences for each compound regardless of dosing regimen and after dose equivalence normalization. The majority of participants in both dosing groups had concentrations expected to elicit clinical response to ETI therapy. These findings indicate that dose reduction may be a viable strategy to maintain clinical benefit while managing intolerance.
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Walmart is a major player in the US retail sector and was one of the grocery corporations that bucked the trend of declining retail sales at the start of the COVID-19 pandemic in 2020. Particularly in the initial stages of the pandemic, governance priorities focussed on restricting the movement of people and closing non-essential retailers and service providers to slow the spread of the virus and keep people safe. This paper investigates the impact of non-pharmaceutical interventions, in the form of lockdown stringency measures, on consumer purchasing behaviours for essential goods over the onset of the pandemic. Focussing on both instore and online sales outcomes for Walmart in the US, we examine changes between pre-pandemic trends in two different sales outcomes, sales transactions and total spend, and trends in 2020. We then employ a series of multi-level regression models to estimate the impact that imposed stringency measures had on these sales outcomes, at both national and state level. Results indicate that nationally consumers were making fewer, larger physical shopping trips and huge increases in online sales was seen ubiquitously across the country. Novel and expansive insights from such a wide-spread retailer, such as Walmart, can help retailers, stakeholders and policy makers understand changing consumption trends to inform business strategies and resilience planning for the future. Furthermore, this study highlighted the value of examining spatial trends in sales outcomes and hopes to influence greater consideration of this in future research.
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Myasthenia gravis is an autoimmune disorder defined by muscle weakness and fatigability associated with antibodies against proteins of the neuromuscular junction (NMJ). The most common autoantibody target is the acetylcholine receptor (AChR). Three mechanisms have been postulated by which autoantibodies might interfere with neurotransmission: direct antagonism of the receptor, complement-mediated destruction of the postsynaptic membrane, and enhanced internalization of the receptor. It is very likely that more than one of these mechanisms act in parallel. Dissecting the mechanisms of autoantibody-mediated pathology requires patient-derived, monoclonal antibodies. Using membrane antigen capture activated cell sorting (MACACS), we isolated AChR-specific B cells from patients with myasthenia gravis, and produced six recombinant antibodies. All AChR-specific antibodies were hypermutated, including isotypes IgG1, IgG3, and IgG4, and recognized different subunits of the AChR. Despite clear binding, none of the individual antibodies showed significant antagonism of the AChR measured in an in vitro neuromuscular synapse model, or AChR-dependent complement activation, and they did not induce myasthenic signs in vivo. However, combinations of antibodies induced strong complement activation in vitro, and severe weakness in a passive transfer myasthenia gravis rat model, associated with NMJ destruction and complement activation in muscle. The strongest complement activation was mediated by combinations of antibodies targeting disparate subunits of the AChR, and such combinations also induced the formation of large clusters of AChR on the surface of live cells in vitro. We propose that synergy between antibodies of different epitope specificities is a fundamental feature of this disease, and possibly a general feature of complement-mediated autoimmune diseases. The importance of synergistic interaction between antibodies targeting different subunits of the receptor can explain the well-known discrepancy between serum anti-AChR titers and clinical severity, and has implications for therapeutic strategies currently under investigation.
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Miastenia Gravis , Animais , Anticorpos Monoclonais , Autoanticorpos , Análise por Conglomerados , Ativação do Complemento , Proteínas do Sistema Complemento , Epitopos , Imunoglobulina G/metabolismo , Miastenia Gravis/patologia , Ratos , Receptores Colinérgicos , Receptores de ComplementoRESUMO
Dengue fever can be caused by one of four distinct dengue virus (DENV) serotypes that cocirculate in many parts of the world. Point of care serotype-specific nonstructural protein-1 (NS1) capture assays for the rapid serotyping of DENV in human sera would greatly support epidemiological surveillance and potentially also prognosis in individual patients. To ensure both serotype specificity and broad coverage of variants within serotypes, we have applied an innovative approach for the generation and selection of serotype-specific anti-NS1 mAbs. To elicit mAbs against conformational epitopes, NMRI mice were immunized with living HEK 293 transfectants expressing the native target Ags in multiple display on the cell surface. For each serotype, three different NS1 sequence variants were sequentially used for immunization of mice, hybridoma selection, and capture assay development, respectively. Selection of optimal combinations of capturing and detecting mAbs yielded highly sensitive and specific NS1 serotyping ELISAs (st-ELISAs) for the four serotypes. st-ELISA testing of 41 dengue patient sera showed a 100% concordance with the serotype determined by serotype-specific reverse transcriptase real-time quantitative PCR. The respective NS1 variants could be detected for â¼10 d after the onset of illness. Ab-dependent enhancement of DENV infections may be associated with a specific range of pre-existing anti-DENV serological Ab titers. Testing of patient sera with the developed st-ELISAs will not only be useful for epidemiological studies and surveillance, but it may also help to develop and validate assays that can distinguish protective versus enhancing Ab responses for risk assessment for the development of severe dengue disease in individual patients.
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Vírus da Dengue/imunologia , Sorotipagem/métodos , Soro/imunologia , Soro/virologia , Proteínas não Estruturais Virais/imunologia , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Linhagem Celular , Reações Cruzadas/imunologia , Dengue/sangue , Dengue/imunologia , Dengue/virologia , Epitopos/imunologia , Células HEK293 , Humanos , Imunização/métodos , Sensibilidade e Especificidade , SorogrupoRESUMO
Occupational therapy practitioners generally agree that sex is a meaningful occupation and should be part of the rehabilitation process; however, there has been reluctance to include clients' sexual concerns in practice. To explore how occupational therapy is addressing clients' sexual concerns, we (one author with a professional background in occupational therapy and the other who is a consumer of occupational therapy services) used a coconstructed autoethnographic approach that involved shared reflection between us. We discuss insights discovered through the coconstructed process, including the potential health risks involved in sexual activity, the importance of sexuality as a core aspect of identity, the ways that rehabilitation currently excludes sexuality, and the potential role of occupational therapy in addressing sexuality.
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Atitude do Pessoal de Saúde , Terapia Ocupacional , Comportamento Sexual , Sexualidade , Antropologia Cultural , Humanos , Acidente Vascular Cerebral , Reabilitação do Acidente Vascular CerebralRESUMO
Symptoms of depression negatively impact on mother-infant relationships and child outcomes. We evaluated a novel, 10-session mother-infant therapeutic playgroup-Community HUGS (CHUGS)-which combines cognitive and experiential components through psychoeducation, play, music, and movement. Participants were mothers experiencing a range of postnatal mental health difficulties, including depression, with infants ≤12 months of age. However, the aim was not to treat maternal depression but to ameliorate associated problems in the mother-infant interaction. In the feasibility study, all participants received CHUGS. In the pilot randomized controlled trial (RCT), participants were randomized between intervention and a wait-list. Outcomes were the Parenting Stress Index (PSI; R.R. Abidin, 1995), Parenting Sense of Competency Scale (Self-Efficacy subscale; J. Gibaud-Wallston & L.P. Wandersman, 1978), and the Depression, Anxiety, Stress Scales (P.F. Lovibond & S.H. Lovibond, 1995). In the feasibility study (n = 74), PSI scores dropped on all subscales, all ps < .01. Depression, p < .001, anxiety, p = .01, stress, p = .01, and self-efficacy, p < .001, all showed improvements, as did observer-rated mother-infant interactions, p < .001. In the RCT, depression, p < .001, anxiety, p = .005, and stress, p < .001, symptoms were significantly reduced for intervention participants (n = 16), as compared to wait-list participants (n = 15). The CHUGS program had high participant satisfaction and produced improvements in self-efficacy, depression, anxiety, stress, and mother-infant interactions that supported the program's acceptability and the utility of further rollout.
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Ansiedade/terapia , Depressão/terapia , Relações Mãe-Filho , Mães , Psicoterapia/métodos , Autoeficácia , Estresse Psicológico/terapia , Adolescente , Adulto , Filho de Pais com Deficiência/psicologia , Estudos de Viabilidade , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Relações Mãe-Filho/psicologia , Mães/psicologia , Avaliação de Resultados em Cuidados de Saúde , Projetos Piloto , Ludoterapia/métodos , Adulto JovemRESUMO
BACKGROUND: Due to their rising incidence and progressive geographical spread, infections with mosquito-borne viruses, such as dengue (DENV), chikungunya and zika virus, have developed into major public health challenges. Since all of these viruses may cause similar symptoms and can occur in concurrent epidemics, tools for their differential diagnosis and epidemiological monitoring are of urgent need. RESULTS: Here we report the application of a novel strategy to rapidly generate monoclonal antibodies (mAbs) against native viral antigens, exemplified for the DENV nonstructural glycoprotein 1 (NS1). The described system is based on the immunization of mice with transfected mammalian cells expressing the target antigens in multiple displays on their cell surface and thereby presenting them efficiently to the host immune system in their native conformation. By applying this cell-based approach to the DENV NS1 protein of serotypes 1 (D1NS1) and 4 (D4NS1), we were able to rapidly generate panels of DENV NS1 serotype cross-reactive, as well as D1NS1- and D4NS1 serotype-specific mAbs. Our data show that the generated mAbs were capable of recognizing the endogenous NS1 protein in DENV-containing biological samples. CONCLUSION: The use of this novel immunization strategy, allows for a fast and efficient generation of hybridoma cell lines, producing mAbs against native viral antigens. Envisaged applications of the mAbs include the development of test platforms enabling a differentiation of the DENV serotypes and high resolution immunotyping for epidemiological studies.
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Anticorpos Monoclonais/imunologia , Células Apresentadoras de Antígenos/imunologia , Antígenos Virais/imunologia , Avaliação Pré-Clínica de Medicamentos/métodos , Imunização/métodos , Proteínas não Estruturais Virais/imunologia , Animais , Desenho de Fármacos , Mapeamento de Epitopos , Células HEK293 , Humanos , Imunoensaio/métodos , CamundongosRESUMO
Rates of child exposure to inter-parental conflict are high and appear to be increasing, with at least one million Australian children affected annually. To-date, there is no established prevalence for inter-parental conflict that includes the more common but less severe forms for young families in the wider Australian community. The current study aims were to examine the prevalence, persistence, and the individual, family and socio-economic context of inter-parental conflict. Data were from four waves of the Baby (n = 4,898) and Kindergarten (n = 4,182) cohorts of the Longitudinal Study of Australian Children. Verbal and physical inter-parental conflict was measured using the Argumentative Relationship Scale. More than 1 in 3 mothers (35-36%) reported any verbal and/or physical conflict. Prevalence of verbal conflict at each wave (10-13%) was higher than physical conflict (4-10%), with low co-occurrence (1-3%). Report of inter-parental conflict at one wave only was most common (19%); although 13% of mothers reported conflict at two waves; and 5-6% reported persistent conflict across three or more waves. Social disadvantage was consistently associated with report at one-wave only and persistent inter-parental conflict. Extrapolated to the Australian population, an estimated 1.9 million children are likely to be affected by inter-parental conflict within any 6 years of the early-to-middle childhood period. Establishing accurate prevalence and understanding the social context of the less severe but most common forms of inter-parental conflict will allow family and child support services to allocate finite resources more effectively and develop targeted interventions to promote children's positive development.
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Desenvolvimento Infantil , Conflito Familiar/psicologia , Pais , Estresse Psicológico/complicações , Austrália/epidemiologia , Criança , Pré-Escolar , Estudos de Coortes , Humanos , Lactente , Estudos Longitudinais , Fatores Socioeconômicos , Estresse Psicológico/etiologia , Inquéritos e QuestionáriosRESUMO
Newborn screening for cystic fibrosis (CF) occasionally results in an inconclusive diagnosis of this disease, and these individuals are designated as CFTR-related metabolic syndrome (CRMS) in the United States, and CF Screen Positive Inconclusive Diagnosis (CFSPID) in other countries. Some of these asymptomatic individuals will progress to symptomatic disease, but risk factors associated with disease progression are not well understood. This scoping review was conducted to comprehensively map nonbiological risk factors in the CRMS/CFSPID literature and to identify understudied topics. Six electronic databases were systematically searched, resulting in 2951 studies. Forty nine eligible works were identified as including information on nonbiological risk factors related to CRMS/CFPSID. Eligible studies were published from 2002 to 2024, most prevalently in the United States (36.7%), and as quantitative data (81.6%). Of the 49 eligible works, 23 articles contributed only intellectual conjecture, while 26 articles contained original data, which underwent full-text qualitative content analysis. Key themes identified in descending order of content coverage included Psychological Impact, Management Care, Newborn Screening and Diagnostics, Communicating Diagnosis, and Lifestyle and External Exposures. This scoping review identified that while nonbiological risk factors are being studied in the CRMS/CFSPID literature, there was nearly equal distribution of works gathering original data to those citing previously published information. These findings indicate a critical need for original data collection on these risk factors, particularly on understudied topics identified herein.
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PURPOSE: To assess the diagnostic performance of distributed human intelligence for the classification of polyp candidates identified with computer-aided detection (CAD) for computed tomographic (CT) colonography. MATERIALS AND METHODS: This study was approved by the institutional Office of Human Subjects Research. The requirement for informed consent was waived for this HIPAA-compliant study. CT images from 24 patients, each with at least one polyp of 6 mm or larger, were analyzed by using CAD software to identify 268 polyp candidates. Twenty knowledge workers (KWs) from a crowdsourcing platform labeled each polyp candidate as a true or false polyp. Two trials involving 228 KWs were conducted to assess reproducibility. Performance was assessed by comparing the area under the receiver operating characteristic curve (AUC) of KWs with the AUC of CAD for polyp classification. RESULTS: The detection-level AUC for KWs was 0.845 ± 0.045 (standard error) in trial 1 and 0.855 ± 0.044 in trial 2. These were not significantly different from the AUC for CAD, which was 0.859 ± 0.043. When polyp candidates were stratified by difficulty, KWs performed better than CAD on easy detections; AUCs were 0.951 ± 0.032 in trial 1, 0.966 ± 0.027 in trial 2, and 0.877 ± 0.048 for CAD (P = .039 for trial 2). KWs who participated in both trials showed a significant improvement in performance going from trial 1 to trial 2; AUCs were 0.759 ± 0.052 in trial 1 and 0.839 ± 0.046 in trial 2 (P = .041). CONCLUSION: The performance of distributed human intelligence is not significantly different from that of CAD for colonic polyp classification.
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Pólipos do Colo/diagnóstico por imagem , Colonografia Tomográfica Computadorizada/métodos , Internet , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Idoso , Algoritmos , Área Sob a Curva , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Curva ROC , Reprodutibilidade dos Testes , Estudos Retrospectivos , Software , Estatísticas não ParamétricasRESUMO
Determining antigen specificity is vital for understanding B cell biology and for producing human monoclonal antibodies. We describe here a powerful method for identifying B cells that recognize membrane antigens expressed on cells. The technique depends on two characteristics of the interaction between a B cell and an antigen-expressing cell: antigen-receptor-mediated extraction of antigen from the membrane of the target cell, and B cell activation. We developed the method using influenza hemagglutinin as a model viral membrane antigen, and tested it using acetylcholine receptor (AChR) as a model membrane autoantigen. The technique involves co-culturing B cells with adherent, bioorthogonally labeled cells expressing GFP-tagged antigen, and sorting GFP-capturing, newly activated B cells. Hemagglutinin-specific B cells isolated this way from vaccinated human donors expressed elevated CD20, CD27, CD71, and CD11c, and reduced CD21, and their secreted antibodies blocked hemagglutination and neutralized viral infection. Antibodies cloned from AChR-capturing B cells derived from patients with myasthenia gravis bound specifically to the receptor on cell membrane. The approach is sensitive enough to detect antigen-specific B cells at steady state, and can be adapted for any membrane antigen.
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Antígenos de Superfície/imunologia , Linfócitos B/imunologia , Separação Celular/métodos , Adulto , Idoso , Animais , Antígenos de Superfície/isolamento & purificação , Autoantígenos/imunologia , Autoantígenos/isolamento & purificação , Subpopulações de Linfócitos B/imunologia , Linhagem Celular Tumoral , Células Clonais , Epitopos de Linfócito B/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Imunofenotipagem , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Miastenia Gravis/imunologia , Receptores Colinérgicos/imunologiaRESUMO
When in public places, many individuals with physical disabilities experience staring. Although staring is typically seen as uncomplicated and something to be "ignored," it has consequences for the person being stared and the staree. Few studies have focused on staring experienced by men following spinal cord injury (SCI). Accordingly, this study explored how adult men with SCI give meaning to the staring from others, the consequences for them, and their responses to the staring and to the starer. Principles of modified grounded theory methods were used to conduct a secondary analysis of interview data for 30 male participants from a larger study of community-dwelling individuals with SCI. Themes revealed through analysis related to context-dependent meanings of staring, negative consequences of staring for some men, and positive opportunities for self-growth and interaction with the public. These findings contribute to a more complex understanding of staring and the relationship between the starer and staree in various social circumstances which can support people living with differences in their public interactions, and improve their quality of life.
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Pessoas com Deficiência/psicologia , Ajustamento Social , Traumatismos da Medula Espinal , Adolescente , Adulto , Humanos , Masculino , Pesquisa Qualitativa , Qualidade de Vida , Traumatismos da Medula Espinal/reabilitação , Adulto JovemRESUMO
BACKGROUND: Much lore but few studies describe a relation of chocolate to mood. We examined the cross-sectional relationship of chocolate consumption with depressed mood in adult men and women. METHODS: A sample of 1018 adults (694 men and 324 women) from San Diego, California, without diabetes or known coronary artery disease was studied in a cross-sectional analysis. The 931 subjects who were not using antidepressant medications and provided chocolate consumption information were the focus of the analysis. Mood was assessed using the Center for Epidemiologic Studies Depression Scale (CES-D). Cut points signaling a positive depression screen result (CES-D score, >or=16) and probable major depression (CES-D score, >or=22) were used. Chocolate servings per week were provided by 1009 subjects. Chocolate consumption frequency and rate data from the Fred Hutchinson Food Frequency Questionnaire were also available for 839 subjects. Chocolate consumption was compared for those with lower vs higher CES-D scores. In addition, a test of trend was performed. RESULTS: Those screening positive for possible depression (CES-D score >or=16) had higher chocolate consumption (8.4 servings per month) than those not screening positive (5.4 servings per month) (P = .004); those with still higher CES-D scores (>or=22) had still higher chocolate consumption (11.8 servings per month) (P value for trend, <.01). These associations extended to both men and women. These findings did not appear to be explained by a general increase in fat, carbohydrate, or energy intake. CONCLUSION: Higher CES-D depression scores were associated with greater chocolate consumption. Whether there is a causal connection, and if so in which direction, is a matter for future prospective study.
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Cacau , Depressão/diagnóstico , Depressão/psicologia , Comportamento Alimentar/psicologia , Adulto , Afeto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Alimentos , Humanos , Masculino , Pessoa de Meia-Idade , Probabilidade , Escalas de Graduação Psiquiátrica , Valores de Referência , Fatores Sexuais , Inquéritos e Questionários , Adulto JovemRESUMO
Prolonged stimulation leads to specific and stable changes in an animal's behavior. In interneurons, this plasticity requires spatial and temporal control of neuronal protein synthesis. Whether such translational control occurs in sensory neurons is not known. Adaptation of the AWC olfactory sensory neurons of C. elegans requires the cGMP-dependent protein kinase EGL-4. Here, we show that the RNA-binding PUF protein FBF-1 is required in the adult AWC for adaptation. In the odor-adapted animal, it increases translation via binding to the egl-4 3' UTR. Further, the PUF protein may localize translation near the sensory cilia and cell body. Although the RNA-binding PUF proteins have been shown to promote plasticity in development by temporally and spatially repressing translation, this work reveals that in the adult nervous system, they can work in a different way to promote experience-dependent plasticity by activating translation in response to environmental stimulation.