RESUMEN
Most cellular ubiquitin signaling is initiated by UBA1, which activates and transfers ubiquitin to tens of E2 enzymes. Clonally acquired UBA1 missense mutations cause an inflammatory-hematologic overlap disease called VEXAS (vacuoles, E1, X-linked, autoinflammatory, somatic) syndrome. Despite extensive clinical investigation into this lethal disease, little is known about the underlying molecular mechanisms. Here, by dissecting VEXAS-causing UBA1 mutations, we discovered that p.Met41 mutations alter cytoplasmic isoform expression, whereas other mutations reduce catalytic activity of nuclear and cytoplasmic isoforms by diverse mechanisms, including aberrant oxyester formation. Strikingly, non-p.Met41 mutations most prominently affect transthioesterification, revealing ubiquitin transfer to cytoplasmic E2 enzymes as a shared property of pathogenesis amongst different VEXAS syndrome genotypes. A similar E2 charging bottleneck exists in some lung cancer-associated UBA1 mutations, but not in spinal muscular atrophy-causing UBA1 mutations, which instead, render UBA1 thermolabile. Collectively, our results highlight the precision of conformational changes required for faithful ubiquitin transfer, define distinct and shared mechanisms of UBA1 inactivation in diverse diseases, and suggest that specific E1-E2 modules control different aspects of tissue differentiation and maintenance.
Asunto(s)
Enzimas Activadoras de Ubiquitina , Enzimas Activadoras de Ubiquitina/metabolismo , Enzimas Activadoras de Ubiquitina/genética , Humanos , Mutación Missense , Ubiquitina/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/metabolismoRESUMEN
An influx of laws permitting recreational cannabis sales in the US has allowed for increased advertising. The purpose of this study was to describe how adolescents perceive outdoor and print cannabis advertising and to identify aspects of advertising that are appealing or eye-catching, focusing on five themes: price promotion, sex appeal, wellness, party, and text-only. We recruited adolescents ages 11-17 to participate in seven focus groups (44 participants) from 2019 to 2020. Participants viewed cannabis advertisements and responses were summarized using deductive thematic analysis. The party-themed advertisements were the most salient to participants, who desired to emulate the behaviors shown. Participants favored ads featuring young people and containing multiple colors. Participants disliked advertisements perceived to portray misleading or contradictory messages, such as the promotion of physical activity or use of sex appeal, and ads perceived to lack authenticity. Identification of youth appealing features can help inform cannabis advertising regulations.
RESUMEN
BACKGROUND: The American College of Chest Physicians (ACCP) and National Institutes for health and care Excellence in the United Kingdom (NICE) recommend that patients who are high risk for thrombotic events but require cessation of oral anticoagulation with warfarin, due to bleeding risk of a planned procedure, undergo bridging therapy with heparin. However, those conditions which are considered high risk are not universal, nor do guidelines differentiate between low molecular weight heparin (LMWH) and unfractionated heparin. Triple positive antiphospholipid syndrome (APS) is a thrombophilic state with a very high risk for thrombotic events during periods of anticoagulation cessation. Patients with secondary antiphospholipid syndrome in the setting of SLE may be at an even greater risk of thrombotic events during the perioperative period. PURPOSE: Along with a review of the literature for perioperative management in APS we present three cases of triple positive secondary APS in systemic lupus erythematosus (SLE) patients who had severe thrombotic complications after cessation of their oral anticoagulation despite being bridged with LWMH. CONCLUSION: Given the severity and rapidity of thrombotic complications with low molecular weight heparin bridging, we propose that all patients with triple positive APS, especially secondary APS with SLE should undergo bridging therapy with intravenous UFH to reduce time without anticoagulation and minimize risk of thrombotic complications. Furthermore, we propose that NICE include APS in the list of medical conditions which are high risk for thrombotic complications and require bridging therapy.
Asunto(s)
Síndrome Antifosfolípido , Lupus Eritematoso Sistémico , Trombosis , Anticoagulantes/efectos adversos , Síndrome Antifosfolípido/complicaciones , Síndrome Antifosfolípido/tratamiento farmacológico , Heparina/efectos adversos , Heparina de Bajo-Peso-Molecular , Humanos , Lupus Eritematoso Sistémico/complicaciones , Lupus Eritematoso Sistémico/tratamiento farmacológico , Trombosis/etiología , Trombosis/prevención & controlRESUMEN
Medically complex children including infants undergoing cardiac surgery are at increased risk for hospital readmissions. Investigation of this population may reveal opportunities to optimize systems and coordination of care. A retrospective study of all infants undergoing cardiac surgery from 2015 through 2016 at a large tertiary institution who were readmitted within 1 year of discharge from cardiac surgical hospitalization was performed. Data specific to patient characteristics, surgical hospitalization, and readmission hospitalization are described. Unplanned readmissions within 1 year of hospital discharge were analyzed with Cox proportional hazard regression to identify factors associated with increased hazard for earlier unplanned readmission. Comparable to previous reports, 12% (78/658) of all surgical hospitalizations were associated with unplanned readmission within 30 days. Infectious etiology, followed by cardiac and gastrointestinal problems, was the most common reasons for unplanned 30-day readmission. Unplanned readmissions within 2 weeks of discharge were multifactorial and less commonly related to cardiac or surgical care. Primary nasogastric tube feeding at the time of discharge was the only significant risk factor for earlier unplanned readmission (p = 0.032) on multivariable analysis. Increased care coordination with particular attention to feeding and comorbidity management may be future targets to effectively mitigate readmissions and improve quality of care in this population.
Asunto(s)
Procedimientos Quirúrgicos Cardíacos/métodos , Cardiopatías Congénitas/cirugía , Readmisión del Paciente/estadística & datos numéricos , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Alta del Paciente/estadística & datos numéricos , Periodo Posoperatorio , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Populations of boreal and southern mountain caribou in Alberta, Canada, are declining, and the ultimate cause of their decline is believed to be anthropogenic disturbance. Linear features are pervasive across the landscape, and of particular importance, seismic lines established in the 1900s (legacy seismic lines) are slow to regenerate. Off-highway vehicles are widely used on these seismic lines and can hamper vegetative re-growth because of ongoing physical damage, compaction, and active clearing. Restoration of seismic lines within caribou range is therefore a priority for the recovery of threatened populations in Alberta, but a triage-type approach is necessary to prioritize restoration and ensure conservation resources are wisely spent. To target restoration efforts, our objective was to determine factors that best explained levels of off-highway vehicles use on seismic lines intersecting roads. We investigated the relative importance of local topography, vegetation attributes of seismic lines, and broad-scale human factors such as the density of infrastructures and the proximity to recreation campsites and towns to explain the observed levels of off-highway vehicles use. We found that off-highway vehicles use was mainly associated with local topography and vegetation attributes of seismic lines that facilitated ease-of-travel. Broad-scale landscape attributes associated with industrial, recreation access, or hunting activities did not explain levels of off-highway vehicles use. Management actions aimed at promoting natural regeneration and reduce ease-of-travel on legacy seismic lines within caribou ranges can be beneficial to caribou recovery in Alberta, Canada, and we therefore recommend restrictions of off-highway vehicles use on low vegetation, dry seismic lines in caribou ranges.
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Conservación de los Recursos Naturales/métodos , Ecosistema , Fenómenos Geológicos , Vehículos a Motor Todoterreno , Reno/crecimiento & desarrollo , Transportes , Alberta , Animales , Industria Procesadora y de Extracción , Bosques , Densidad de Población , Recreación , Estaciones del AñoRESUMEN
OBJECTIVE: Tissue damage in lupus nephritis (LN) is mediated by activation of the classical complement pathway. Complement-mediated upregulation of endothelial cell adhesion molecules is seen in dermal blood vessels of non-lesional skin of patients with active lupus. In diseases with systemic complement activation, extensive microvascular C5b-9 deposition is seen in non-lesional skin. In this study, we assess the presence of systemic complement pathway activation as determined by non-lesional skin microvascular C5b-9 deposition in patients with LN. METHODS: Eight patients with active LN and eight patients without active LN underwent non-lesional skin biopsies. Using a diaminobenzidine technique, specimens were evaluated for microvascular C5b-9 consistent with systemic complement pathway activation. RESULTS: Five of eight patients with active LN and one of eight patients without active LN demonstrated positive C5b-9 staining in non-lesional skin (p=0.04). Positive non-lesional C5b-9 staining has greater specificity, 87.5%, for active LN than pyuria, low complements, elevated double-stranded DNA (dsDNA) and proteinuria. Urine protein creatinine ratio was significantly higher in patients with positive non-lesional C5b-9 deposition (5.18 vs 1.20; p=0.04). C5b-9 deposition was not associated with a higher NIH Activity Index, interstitial fibrosis, dsDNA or lower complements. CONCLUSION: This is the first study to demonstrate evidence in non-lesional skin of microvascular C5b-9 indicative of systemic complement pathway activation in LN. C5b-9 deposition is statistically more common and demonstrated greater specificity than most historical biomarkers for active LN. The findings support a potential role for microvascular C5b-9 assessment in non-lesional skin as a biomarker for LN activity.
Asunto(s)
Lupus Eritematoso Sistémico , Nefritis Lúpica , Humanos , Nefritis Lúpica/complicaciones , Nefritis Lúpica/patología , Complejo de Ataque a Membrana del Sistema Complemento , Proteínas del Sistema Complemento , Activación de Complemento , BiomarcadoresRESUMEN
Most cellular ubiquitin signaling is initiated by UBA1, which activates and transfers ubiquitin to tens of E2 enzymes. Clonally acquired UBA1 missense mutations cause an inflammatory-hematologic overlap disease called VEXAS (vacuoles, E1, X-linked, autoinflammatory, somatic) syndrome. Despite extensive clinical investigation into this lethal disease, little is known about the underlying molecular mechanisms. Here, by dissecting VEXAS-causing UBA1 mutations, we discovered that p.Met41 mutations alter cytoplasmic isoform expression, whereas other mutations reduce catalytic activity of nuclear and cytoplasmic isoforms by diverse mechanisms, including aberrant oxyester formation. Strikingly, non-p.Met41 mutations most prominently affect transthioesterification, revealing ubiquitin transfer to cytoplasmic E2 enzymes as a shared property of pathogenesis amongst different VEXAS syndrome genotypes. A similar E2 charging bottleneck exists in some lung cancer-associated UBA1 mutations, but not in spinal muscular atrophy-causing UBA1 mutations, which instead, render UBA1 thermolabile. Collectively, our results highlight the precision of conformational changes required for faithful ubiquitin transfer, define distinct and shared mechanisms of UBA1 inactivation in diverse diseases, and suggest that specific E1-E2 modules control different aspects of tissue differentiation and maintenance.
RESUMEN
A 31-year-old male presented to our facility with complaints of shortness of breath and left-sided chest pain. On record review, it was revealed that he had been seen in 2014 for an almost identical presentation and had been found to have haemolytic anaemia with warm autoantibodies. Following his acute treatment during that hospital admission, he was lost to follow-up. During his subsequent admission, 5 years later, he was found to have a systemic autoimmune disorder with a superimposed acute bacterial infection leading to a second case of haemolytic anaemia and at this time with both cold and warm antibodies present. While his diagnosis was initially difficult to make due to both derangements in expected laboratory values and the mixed pattern of the haemolytic anaemia, he was promptly treated with intravenous immune globulin and steroids and was able to make a full recovery.
Asunto(s)
Anemia Hemolítica Autoinmune/inmunología , Autoanticuerpos/sangre , Lupus Eritematoso Sistémico/complicaciones , Adulto , Anemia Hemolítica Autoinmune/diagnóstico , Anemia Hemolítica Autoinmune/tratamiento farmacológico , Crioglobulinas/análisis , Diagnóstico Diferencial , Disnea/etiología , Glucocorticoides/uso terapéutico , Humanos , Inmunoglobulinas Intravenosas/uso terapéutico , MasculinoRESUMEN
Analyte isolation is an important process that spans a range of biomedical disciplines, including diagnostics, research, and forensics. While downstream analytical techniques have advanced in terms of both capability and throughput, analyte isolation technology has lagged behind, increasingly becoming the bottleneck in these processes. Thus, there exists a need for simple, fast, and easy to integrate analyte separation protocols to alleviate this bottleneck. Recently, a new class of technologies has emerged that leverages the movement of paramagnetic particle (PMP)-bound analytes through phase barriers to achieve a high efficiency separation in a single or a few steps. Specifically, the passage of a PMP/analyte aggregate through a phase interface (aqueous/air in this case) acts to efficiently "exclude" unbound (contaminant) material from PMP-bound analytes with higher efficiency than traditional washing-based solid-phase extraction (SPE) protocols (i.e., bind, wash several times, elute). Here, we describe for the first time a new type of "exclusion-based" sample preparation, which we term "AirJump". Upon realizing that much of the contaminant carryover stems from interactions with the sample vessel surface (e.g., pipetting residue, wetting), we aim to eliminate the influence of that factor. Thus, AirJump isolates PMP-bound analyte by "jumping" analyte directly out of a free liquid/air interface. Through careful characterization, we have demonstrated the validity of AirJump isolation through comparison to traditional washing-based isolations. Additionally, we have confirmed the suitability of AirJump in three important independent biological isolations, including protein immunoprecipitation, viral RNA isolation, and cell culture gene expression analysis. Taken together, these data sets demonstrate that AirJump performs efficiently, with high analyte yield, high purity, no cross contamination, rapid time-to-isolation, and excellent reproducibility.
Asunto(s)
Extracción en Fase Sólida/métodos , Perfilación de la Expresión Génica , Inmunoprecipitación , Proteínas/aislamiento & purificación , ARN Viral/aislamiento & purificación , Reproducibilidad de los ResultadosRESUMEN
The monitoring of viral load is critical for proper management of antiretroviral therapy for HIV-positive patients. Unfortunately, in the developing world, significant economic and geographical barriers exist, limiting access to this test. The complexity of current viral load assays makes them expensive and their access limited to advanced facilities. We attempted to address these limitations by replacing conventional RNA extraction, one of the essential processes in viral load quantitation, with a simplified technique known as immiscible filtration assisted by surface tension (IFAST). Furthermore, these devices were produced via the embossing of wax, enabling local populations to produce and dispose of their own devices with minimal training or infrastructure, potentially reducing the total assay cost. In addition, IFAST can be used to reduce cold chain dependence during transportation. Viral RNA extracted from raw samples stored at 37°C for 1 week exhibited nearly complete degradation. However, IFAST-purified RNA could be stored at 37°C for 1 week without significant loss. These data suggest that RNA isolated at the point of care (eg, in a rural clinic) via IFAST could be shipped to a central laboratory for quantitative RT-PCR without a cold chain. Using this technology, we have demonstrated accurate and repeatable measurements of viral load on samples with as low as 50 copies per milliliter of sample.
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Filtración/instrumentación , Infecciones por VIH/diagnóstico , VIH/aislamiento & purificación , ARN Viral/aislamiento & purificación , Secuencia de Bases , Diseño de Equipo , Filtración/métodos , VIH/genética , Infecciones por VIH/virología , Humanos , Límite de Detección , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Tensión Superficial , Carga ViralRESUMEN
While potentially powerful, access to molecular diagnostics is substantially limited in the developing world. Here we present an approach to reduced cost molecular diagnostic instrumentation that has the potential to empower developing world communities by reducing costs through streamlining the sample preparation process. In addition, this instrument is capable of producing its own consumable devices on demand, reducing reliance on assay suppliers. Furthermore, this instrument is designed with an "open" architecture, allowing users to visually observe the assay process and make modifications as necessary (as opposed to traditional "black box" systems). This open environment enables integration of microfluidic fabrication and viral RNA purification onto an easy-to-use modular system via the use of interchangeable trays. Here we employ this system to develop a protocol to fabricate microfluidic devices and then use these devices to isolate viral RNA from serum for the measurement of human immunodeficiency virus (HIV) viral load. Results obtained from this method show significantly reduced error compared with similar nonautomated sample preparation processes.
Asunto(s)
Métodos Analíticos de la Preparación de la Muestra/instrumentación , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas de Diagnóstico Molecular/instrumentación , ARN Viral/análisis , Robótica/instrumentación , Métodos Analíticos de la Preparación de la Muestra/economía , Benchmarking , Errores Diagnósticos/prevención & control , VIH/aislamiento & purificación , VIH/metabolismo , Infecciones por VIH/sangre , Infecciones por VIH/diagnóstico , Infecciones por VIH/economía , Infecciones por VIH/virología , Costos de la Atención en Salud , Humanos , Dispositivos Laboratorio en un Chip/economía , Técnicas Analíticas Microfluídicas/economía , Técnicas de Diagnóstico Molecular/economía , Prueba de Estudio Conceptual , ARN Viral/sangre , ARN Viral/aislamiento & purificación , ARN Viral/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa/economía , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/economía , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/instrumentación , Robótica/economía , Carga Viral , Ceras/químicaRESUMEN
High dose methotrexate has become one of the treatments of choice for patients with primary CNS lymphomas due to its ability to penetrate the blood-brain barrier. A potentially serious complication of this therapy is methotrexate-related nephrotoxicity. We report the case of a patient with a common genetic polymorphism that may have predisposed this patient experience clinically significant toxicity from systemic folate depletion. After the first cycle of chemotherapy that included high dose methotrexate, the patient's serum creatinine rose and the patient's methotrexate level remained above the toxic range for six days. On cycle two, the patient was treated with a 25% dose reduction in methotrexate and more aggressive hydration and alkalization. With this alteration in the regimen, the patient was able to receive six more cycles and had a complete radiographic tumor response in the brain and a disappearance of tumor cells in the CSF without any further renal complications. This case report illustrates the feasibility of administering high dose methotrexate with modifications as a treatment of choice in individuals with methylenetetrahydrofolate reductase gene mutations.