RESUMO
Hidradenitis suppurativa (HS) is a chronic inflammatory skin condition that can manifest with abscesses, sinus tracts, and scarring in the intertriginous areas of the body. HS is characterized by immune dysregulation, featuring elevated levels of myeloid cells, T helper (Th) cells, and pro-inflammatory cytokines, particularly those involved in Th1- and Th17-mediated immunity. In most epidemiological studies, HS shows a strong female sex bias, with reported female-to-male ratios estimated at roughly 3:1, suggesting that sex-related factors contribute to HS pathophysiology. In this article, we review the role of intrinsic and extrinsic factors that contribute to immunological differences between the sexes and postulate their role in the female sex bias observed in HS. We discuss the effects of hormones, X chromosome dosage, genetics, the microbiome, and smoking on sex-related differences in immunity to postulate potential immunological mechanisms in HS pathophysiology. Future studies are required to better characterize sex-biased factors that contribute to HS disease presentations.
Assuntos
Hidradenite Supurativa , Masculino , Humanos , Feminino , Hidradenite Supurativa/etiologia , Sexismo , Citocinas , Células Th17 , AbscessoRESUMO
Perfluorocarbon liquid (PFCL) is an important adjunct in pars plana vitrectomy (PPV) for complex retinal detachment (RD). Complete removal of PFCL is critical to prevent retinal inflammation and cellular toxicity, but removal is not risk-free. We report a case of a new postoperative onset paracentral visual field defect after PPV with PFCL use for treatment of a macula-on RD. We present pre- and postoperative imaging that suggests a likely perioperative iatrogenic cause. [Ophthalmic Surg Lasers Imaging Retina 2022;53:644-646.].
Assuntos
Fluorocarbonos , Descolamento Retiniano , Humanos , Vitrectomia/efeitos adversos , Vitrectomia/métodos , Descolamento Retiniano/diagnóstico , Descolamento Retiniano/cirurgia , Descolamento Retiniano/etiologia , Campos Visuais , Acuidade Visual , Fluorocarbonos/efeitos adversos , Transtornos da Visão/cirurgia , Doença Iatrogênica , Estudos RetrospectivosRESUMO
Cerebral small vessel disease (SVD) is a prevalent disease of aging and a major contributor to stroke and dementia. The most commonly inherited SVD, CADASIL, is caused by dominantly acting cysteine-altering mutations in NOTCH3. These mutations change the number of cysteines from an even to an odd number, but the impact of these alterations on NOTCH3 protein structure remain unclear. Here, we prepared wildtype and four mutant recombinant NOTCH3 protein fragments to analyze the impact of CADASIL mutations on oligomerization, thiol status, and protein stability. Using gel electrophoresis, tandem MS/MS, and collision-induced unfolding, we find that NOTCH3 mutant proteins feature increased amounts of inappropriate disulfide bridges, reduced cysteines, and structural instability. Presence of a second protein factor, an N-terminal fragment of NOTCH3 (NTF), is capable of further altering disulfide statuses of both wildtype and mutant proteins, leading to increased numbers of reduced cysteines and further destabilization of NOTCH3 structure. In sum, these studies identify specific cysteine residues alterations and quaternary structure induced by CADASIL mutations in NOTCH3; further, we validate that reductive factors alter the structure and stability of this small vessel disease protein.
Assuntos
CADASIL , Demência Vascular , Receptor Notch3 , CADASIL/genética , CADASIL/metabolismo , Cisteína/genética , Dissulfetos , Humanos , Proteínas Mutantes , Receptor Notch3/genética , Receptores Notch/metabolismo , Espectrometria de Massas em TandemRESUMO
Cysteine oxidation states of extracellular proteins participate in functional regulation and in disease pathophysiology. In the most common inherited dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), mutations in NOTCH3 that alter extracellular cysteine number have implicated NOTCH3 cysteine states as potential triggers of cerebral vascular smooth muscle cytopathology. In this report, we describe a novel property of the second EGF-like domain of NOTCH3: its capacity to alter the cysteine redox state of the NOTCH3 ectodomain. Synthetic peptides corresponding to this sequence (NOTCH3 N-terminal fragment 2, NTF2) readily reduce NOTCH3 N-terminal ectodomain polypeptides in a dose- and time-dependent fashion. Furthermore, NTF2 preferentially reduces regional domains of NOTCH3 with the highest intensity against EGF-like domains 12-15. This process requires cysteine residues of NTF2 and is also capable of targeting selected extracellular proteins that include TSP2 and CTSH. CADASIL mutations in NOTCH3 increase susceptibility to NTF2-facilitated reduction and to trans-reduction by NOTCH3 produced in cells. Moreover, NTF2 forms complexes with the NOTCH3 ectodomain, and cleaved NOTCH3 co-localizes with the NOTCH3 ectodomain in cerebral arteries of CADASIL patients. The potential for NTF2 to reduce vascular proteins and the enhanced preference for it to trans-reduce mutant NOTCH3 implicate a role for protein trans-reduction in cerebrovascular pathological states such as CADASIL.
Assuntos
CADASIL , Doenças de Pequenos Vasos Cerebrais , CADASIL/genética , CADASIL/metabolismo , Cisteína/genética , Fator de Crescimento Epidérmico/genética , Fator de Crescimento Epidérmico/farmacologia , Humanos , Mutação , Receptor Notch3/genética , Receptor Notch3/metabolismo , Receptores Notch/genética , Receptores Notch/metabolismoRESUMO
Cerebral small vessel disease is a common condition linked to dementia and stroke. As an age-dependent brain pathology, cerebral SVD may share molecular processes with core neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Many neurodegenerative diseases feature abnormal protein accumulation and aberrant protein folding, resulting in multimerization of specific proteins. We investigated if a small NOTCH3 N-terminal fragment (NTF) that co-registers with pathologically affected cells in the inherited SVD, CADASIL, is capable of multimerization. We also characterized endogenous small molecule vascular enhancers and inhibitors of multimerization. NTF multimerizes spontaneously and also forms conjugates with vascular catecholamines, including dopamine and norepinephrine, which avidly promote multimerization of the protein. Inhibition of catecholamine-dependent multimerization by vitamin C and reversal by reducing agents implicate an essential role of oxidation in NTF multimerization. Antibodies that react with degenerating arteries in CADASIL tissue preferentially bind to multimerized forms of NTF. These studies suggest that multimerization of proteins in the aging brain is not restricted to neuronal molecules and may participate in age-dependent vascular pathology.
Assuntos
CADASIL , Catecolaminas/farmacologia , Multimerização Proteica/fisiologia , Receptor Notch3/química , Humanos , Oxirredução , Fragmentos de Peptídeos/química , Compostos de Sulfidrila/químicaRESUMO
The small-vessel disorder cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) arises from mutations in the human gene encoding NOTCH3 and results in vascular smooth muscle cell degeneration, stroke, and dementia. However, the structural changes in NOTCH3 involved in CADASIL etiology are unclear. Here, we discovered site-specific fragmentation of NOTCH3 protein in pathologically affected vessels of human CADASIL-affected brains. EM-based experiments to pinpoint NOTCH3 localization in these brains indicated accumulation of NOTCH3 fragmentation products in the basement membrane, collagen fibers, and granular osmiophilic material within the cerebrovasculature. Using antibodies generated against a disease-linked neo-epitope found in degenerating vascular medium of CADASIL brains, we mapped the site of fragmentation to the NOTCH3 N terminus at the peptide bond joining Asp80 and Pro81 Cleavage at this site was predicted to separate the first epidermal growth factor (EGF)-like domain from the remainder of the protein. We found that the cleavage product from this fragmentation event is released into the conditioned medium of cells expressing recombinant NOTCH3 fragments. Mutagenesis of Pro81 abolished the fragmentation, and low pH and reducing conditions enhanced NOTCH3 proteolysis. Furthermore, substitution of multiple cysteine residues of the NOTCH3 N terminus activated proteolytic release of the first EGF-like repeat, suggesting that the elimination of multiple disulfide bonds in NOTCH3 accelerates its fragmentation. These characteristics link the signature molecular genetic alterations present in individuals with CADASIL to a post-translational protein alteration in degenerating brain arteries. The cellular consequences of these pathological NOTCH3 fragments are an important area for future investigation.
Assuntos
CADASIL/genética , Doenças de Pequenos Vasos Cerebrais/genética , Proteólise , Receptor Notch3/genética , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/patologia , Encéfalo/metabolismo , Encéfalo/patologia , CADASIL/patologia , Doenças de Pequenos Vasos Cerebrais/patologia , Humanos , Mutação/genética , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologiaRESUMO
BACKGROUND: Bariatric surgery patients with mental illness may experience worse surgical outcomes compared to those without. Depression is the most prevalent mental health diagnosis amongst Americans with obesity. Accurate diagnosis and treatment is of paramount importance to mitigate perioperative risk. Unfortunately, there is no standard method to screen patients for depression prior to surgery. Our goal was to understand the relationship between traditional clinical screening tools and a novel patient-reported depression screening survey, Patient Health Questionnaire 8 (PHQ-8), in the setting of the bariatric surgery preoperative assessment. METHODS: The study included all adult bariatric surgery patients from January 2014 through June 2016. Patients who were not assessed using both the PHQ-8 and a traditional clinical depression screening were excluded from the study. There were a total of 4486 patients who met the eligibility criteria and were included in analysis. We used comparative statistics to examine the association between these screening tools and to test for contributing demographic, surgical, and socioeconomic factors. RESULTS: The overall rate of clinically diagnosed depression in the study cohort was 45.6%. In comparison, 14.8% of all patients screened positive for depression using the PHQ-8. Of the patients without a traditional clinical diagnosis of depression, 10.2% screened positive for depression using the PHQ-8. This subset of undiagnosed patients was more likely to be non-white, employed, and had a higher BMI than their clinically diagnosed counterparts. CONCLUSIONS AND RELEVANCE: We found a higher rate of clinically diagnosed depression in our cohort compared to the general population. However, when using the validated PHQ-8 survey, the rate of depression more closely approximated the national incidence. Further, a significant proportion of patients were undiagnosed and/or misdiagnosed by current clinical assessments. Standardizing preoperative depression screening using validated patient-centered tools may prevent the consequences of untreated depression.
Assuntos
Cirurgia Bariátrica , Depressão/diagnóstico , Questionário de Saúde do Paciente , Escalas de Graduação Psiquiátrica , Feminino , Humanos , Masculino , Michigan , Pessoa de Meia-Idade , Cuidados Pré-Operatórios , Sistema de RegistrosRESUMO
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy CADASIL is caused by more than a hundred NOTCH3 mutations. Virtually all encoded mutant proteins contain an odd number of cysteines. As such, structural changes in NOTCH3 may be the primary molecular abnormality in CADASIL. Thus, we sought evidence for structurally altered NOTCH3 protein in CADASIL tissue. Four antibodies were raised in rabbits against two non-overlapping N-terminal NOTCH3 sequences. These reagents were used in immunohistochemical experiments to detect epitopes in post-mortem CADASIL brains (n=8), control brains, and cells overexpressing NOTCH3. To determine the biochemical nature of NOTCH3 epitopes, we used these antibodies to probe pure NOTCH3-Fc fusion proteins treated with acid, urea, guanidinium, ionic detergents, acrylamide, and thiol- and phosphorus-based reductants. All antibodies avidly stained arteries in 8 of 8 CADASIL brain samples. The most prominent staining was in degenerating media of leptomeningeal arteries and sclerotic penetrating vessels. Normal appearing vessels from control brains were not reactive. Antibodies did not react with cultured cells overexpressing NOTCH3 or with purified NOTCH3-Fc protein. Furthermore, treatment of pure protein with acid, chaotropic denaturants, alkylators, and detergents failed to unmask N-terminal NOTCH3 epitopes. Antibodies, however, recognized novel N-terminal epitopes in purified NOTCH3-Fc protein treated with three different reductants (DTT, beta-mercaptoethanol, and TCEP). We conclude that CADASIL arteries feature latent N-terminal NOTCH3 epitopes, suggesting the first evidence in vivo of NOTCH3 structural alterations.