RESUMEN
BACKGROUND: Maternal overweight and obesity have been associated with an increased risk of atopic dermatitis (AD) in the offspring, but the underlying mechanisms are unclear. Vernix caseosa (VC) is a proteolipid material covering the fetus produced during skin development. However, whether maternal prepregnancy weight excess influences fetal skin development is unknown. Characterizing the VC of newborns from mothers with prepregnancy overweight and obesity might reveal AD-prone alterations during fetal skin development. OBJECTIVE: We sought to explore AD biomarkers and staphylococcal loads in VC from the offspring of mothers who were overweight/obese (O/O) before pregnancy versus in those from offspring of normal weight mothers. METHODS: The VC of newborns of 14 O/O and 12 normal weight mothers were collected immediately after birth. Biomarkers were determined by ELISA and staphylococcal species by quantitative PCR. RESULTS: The VC from the O/O group showed decreased expression of skin barrier proteins (filaggrin and loricrin) and increased levels of proinflammatory biomarkers (IgA, thymic stromal lymphopoietin [TSLP], S100A8, IL-25, and IL-33). No differences in concentrations of antimicrobial peptides and enzymes were detected. The VC from the O/O group had a lower Staphylococcus epidermidis and Staphylococcus hominis commensal bacterial load, whereas Staphylococcus aureus bacterial load was not significantly different between the 2 groups. Maternal body mass index was negatively correlated with VC filaggrin expression and S epidermidis load and was positively associated with TSLP concentration. One-year follow-up established that the offspring of O/O mothers had a higher incidence of AD that was specifically linked with decreased VC filaggrin expression and lower S epidermidis load. CONCLUSIONS: VC from neonates of mothers with prepregnancy overweight and obesity exhibit skin barrier molecular alterations and staphylococcal dysbiosis that suggest early mechanistic clues to this population's increased risk of AD.
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Dermatitis Atópica , Obesidad Materna , Vernix Caseosa , Humanos , Recién Nacido , Femenino , Embarazo , Dermatitis Atópica/patología , Proteínas Filagrina , Obesidad Materna/metabolismo , Obesidad Materna/patología , Vernix Caseosa/metabolismo , Sobrepeso , Piel/patología , Citocinas/metabolismo , Linfopoyetina del Estroma Tímico , Obesidad/patología , Biomarcadores/metabolismoRESUMEN
BACKGROUND: Skin tissue engineering is a rapidly evolving field of research that effectively combines stem cells and biological scaffolds to replace damaged tissues. Human Wharton's jelly mesenchymal stromal cells (hWJ-MSCs) are essential to generate tissue constructs, due to their potent immunomodulatory effects and release of paracrine factors for tissue repair. Here, we investigated whether hWJ-MSC grown on human acellular dermal matrix (hADM) scaffolds and exposed to a proinflammatory environment maintain their ability to produce in vitro growth factors involved in skin injury repair and promote in vivo wound healing. METHODS: We developed a novel method involving physicochemical and enzymatic treatment of cadaveric human skin to obtain hADM scaffold. Subsequently, skin bioengineered constructs were generated by seeding hWJ-MSCs on the hADM scaffold (construct 1) and coating it with human platelet lysate clot (hPL) (construct 2). Either construct 1 or 2 were then incubated with proinflammatory cytokines (IL-1α, IL-1ß, IL-6, TNF-α) for 12, 24, 48, 72 and 96 h. Supernatants from treated and untreated constructs and hWJ-MSCs on tissue culture plate (TCP) were collected, and concentration of the following growth factors, bFGF, EGF, HGF, PDGF, VEGF and Angiopoietin-I, was determined by immunoassay. We also asked whether hWJ-MSCs in the construct 1 have potential toward epithelial differentiation after being cultured in an epithelial induction stimulus using an air-liquid system. Immunostaining was used to analyze the synthesis of epithelial markers such as filaggrin, involucrin, plakoglobin and the mesenchymal marker vimentin. Finally, we evaluated the in vivo potential of hADM and construct 1 in a porcine full-thickness excisional wound model. RESULTS: We obtained and characterized the hADM and confirmed the viability of hWJ-MSCs on the scaffold. In both constructs without proinflammatory treatment, we reported high bFGF production. In contrast, the levels of other growth factors were similar to the control (hWJ-MSC/TCP) with or without proinflammatory treatment. Except for PDGF in the stimulated group. These results indicated that the hADM scaffold maintained or enhanced the production of these bioactive molecules by hWJ-MSCs. On the other hand, increased expression of filaggrin, involucrin, and plakoglobin and decreased expression of vimentin were observed in constructs cultured in an air-liquid system. In vivo experiments demonstrated the potential of both hADM and hADM/hWJ-MSCs constructs to repair skin wounds with the formation of stratified epithelium, basement membrane and dermal papillae, improving the appearance of the repaired tissue. CONCLUSIONS: hADM is viable to fabricate a tissue construct with hWJ-MSCs able to promote the in vitro synthesis of growth factors and differentiation of these cells toward epithelial lineage, as well as, promote in a full-thickness skin injury the new tissue formation. These results indicate that hADM 3D architecture and its natural composition improved or maintained the cell function supporting the potential therapeutic use of this matrix or the construct for wound repair and providing an effective tissue engineering strategy for skin repair.
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Dermis Acelular , Células Madre Mesenquimatosas , Gelatina de Wharton , Humanos , Animales , Porcinos , Proteínas Filagrina , Vimentina/metabolismo , Dermis Acelular/metabolismo , gamma Catenina/metabolismo , gamma Catenina/farmacología , Diferenciación Celular , Células Madre Mesenquimatosas/metabolismoRESUMEN
Introdução: Mutações do gene da filagrina vêm sendo associadas, classicamente, a alterações da barreira epitelial em doenças alérgicas com comprometimento da pele e das superfícies mucosas. Particularmente na dermatite atópica, a relação entre filagrina, mecanismo fisiopatológico e evolução clínica tem sido demonstrada. Recentemente, alterações da barreira epitelial com redução da expressão da filagrina, também têm sido associadas a mecanismos imunológicos envolvidos na patogênese da esofagite eosinofílica. Devido a disfunções na barreira epitelial, microrganismos e alérgenos são capazes de penetrarem no epitélio da mucosa esofágica, assim como na dermatite atópica. Objetivo: Avaliar a possível correlação da expressão da filagrina com os achados histopatológicos em biópsias esofágicas de pacientes com esofagite eosinofílica. Métodos: A expressão da filagrina foi investigada in situ, por imuno-histoquímica, em biópsias esofágicas nos seguintes grupos: Grupo I, controle (n=8), amostras provenientes de pacientes saudáveis; Grupo II (n=27), amostras provenientes de pacientes com esofagite eosinofílica. Resultados: Os resultados demonstraram uma diminuição da expressão da filagrina na mucosa do esôfago de portadores de esofagite eosinofílica. Adicionalmente, a intensidade da marcação imuno-histoquímica foi menor na mucosa esofágica com maior infiltração de eosinófilos. Conclusão: A diminuição da expressão de filagrina pode ser um fenomeno fisiopatológico associado ao aumento da quantidade de eosinófilos na mucosa esofágica, podendo impactar na evolução clínica da esofagite eosinofílica.
Introduction:Filaggrin gene mutations have been classically associated with changes in the epithelial barrier in allergic diseases involving the skin and mucosal surfaces. Particularly in atopic dermatitis, the relationship between filaggrin, pathophysiological mechanism and clinical evolution hás been demonstrated. Recently, changes in the epithelial barrier with reduced expression of filaggrin have also been associated with immunological mechanisms involved in the pathogenesis of eosinophilic esophagitis. Due to dysfunction in the epithelial barrier, microorganisms and allergens are able to penetrate the epithelium of the esophageal mucosa, as well as in atopic dermatitis. Objective: To evaluated the possible correlation of filaggrin expression with histopathological findings in esophageal biopsies of patients with eosinophilic esophagitis. Methods: Filaggrin expression was investigated in situ by immunohistochemistry in esophageal biopsies in the following groups: Group I, control (n = 8), samples from healthy patients; Group II (n = 27), samples from patients with eosinophilic esophagitis. Results: The results demonstrated a decrease in the expression of filaggrin in the esophageal mucosa of patients with eosinophilic esophagitis. Additionally, the intensity of the immunohistochemical labeling was lower in the esophageal mucosa with greater infiltration of eosinophils. Conclusion: The reduction of filaggrin expression may be a pathophysiological phenomenon associated with an increase in the quantity of eosinophils in the esophageal mucosa, which may impact on the clinical evolution of eosinophilic esophagitis.
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Humanos , Biopsia , Esofagitis Eosinofílica , Proteínas Filagrina , Pacientes , Piel , Inmunohistoquímica , Alérgenos , Dermatitis Atópica , Mucosa Esofágica , MutaciónRESUMEN
BACKGROUND: Atopic dermatitis (AD) is an itchy, chronic and inflammatory skin condition, with dysfunctional immune response and skin barrier defects. Reduction of filaggrin (FLG) and tight junctions (TJ) proteins, such as claudin-1 (CLDN-1), expression in cutaneous epithelial barrier is remarkable in AD pathogenesis. Ocular involvement occurs in approximately 40% of AD patients leading to changes in the structure of the conjunctiva. OBJECTIVES: We aimed to evaluate the expression of FLG and CLDN-1 in the ocular surface of adults with AD, analysing bulbar conjunctival cells collected by a novel non-invasive cellular imprint. METHODS: Bulbar conjunctival epithelial cells were collected by cellular imprint technique, and FLG and CLDN-1 expression were assessed by immunofluorescence (IF) and real-time polymerase chain reaction (RT-PCR). RESULTS: We detected increased expression of FLG and CLDN-1, as well as their transcript levels in AD patients compared with healthy controls (HC). There was a positive correlation between tear film break-up time (TBUT) and FLG expression. Fluorescein staining was inversely associated with FLG expression. CONCLUSIONS: Our results may reflect a reactive response of the ocular surface to AD-related ocular inflammation and associated dry eye disease. Further investigations focusing on the role of FLG and TJ expression in the ocular surface of AD patients may increment the understanding of the pathophysiology of extracutaneous AD and developing future targeted therapies.
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Dermatitis Atópica , Proteínas Filagrina , Claudina-1/genética , Dermatitis Atópica/genética , Humanos , Proteínas de Filamentos Intermediarios/genética , Proteínas de Filamentos Intermediarios/metabolismo , Mutación , Piel/metabolismoRESUMEN
BACKGROUND: Filaggrin (FLG) loss-of-function variants are major genetic risk factors for atopic dermatitis (AD), but these have not been studied in Latin American populations with and without AD. METHODS: FLG variants R501X and 2282del4 were genotyped in 275 Chilean adults with and without AD from the "Early origins of allergy and asthma" (ARIES) cohort and in 227 patients from an AD cohort based in Santiago, Chile. RESULTS: Among adults in the ARIES cohort, 3.3% were carriers of R501X and 2.9% of 2282del4 variants, all heterozygotes. In this cohort, 6.2% were FLG variant carriers: 11.1% of subjects reporting AD were carriers of FLG variants vs. 5.2% in those without AD (P = 0.13). In this first cohort, FLG variants were not significantly associated with asthma, allergic rhinitis, or food allergy. In the AD cohort, the prevalence of FLG variants was 7% for R501X, 2.2% for the 2282del4 variant, and 9.3% for the combined genotype. In this cohort, FLG variants were present in 15.5% of severe AD vs. 7.1% of mild-to-moderate AD subjects (P = 0.056). Evaluation of Chilean population from both cohorts combined (n = 502) revealed that FLG variants were not significantly associated with AD (OR = 1.92 [95% CI 0.95-3.9], P = 0.067) but were associated with asthma (OR = 2.16 [95% CI 1.02-4.56], P = 0.039). CONCLUSIONS: This is the first study to evaluate FLG loss-of-function variants R501X and 2282del4 in Latin American population, revealing a similar prevalence of these FLG variant carriers to that of European populations. Among Chileans, FLG variants were significantly associated with asthma but not AD.
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Dermatitis Atópica , Proteínas Filagrina/genética , Adulto , Chile/epidemiología , Dermatitis Atópica/epidemiología , Dermatitis Atópica/genética , Predisposición Genética a la Enfermedad , Humanos , Mutación , PrevalenciaRESUMEN
A hidratação cutânea ocorre, em parte, pelos componentes do Fator de Hidratação Natural (NMF), originados da degradação da filagrina, sendo alguns exemplos o ácido pirrolidona-carboxílico (PCA), o ácido urocânico (UCA) e a histidina (His). Estes estão presentes no estrato córneo (EC). O objetivo deste projeto de pesquisa foi determinar, por cromatografia líquida de alta eficiência (CLAE), o PCA, o UCA e a His no estrato córneo de participantes obtido por tape stripping em função do fototipo e idade do participante da pesquisa. Participantes foram selecionados em função da idade acima de 18 anos, ambos os gêneros e fototipo da pele entre I a VI, de acordo com a classificação de Fitzpatrick. As amostras do EC foram obtidas do antebraço volar por tape stripping e irradiadas artificialmente. A cromatografia líquida de alta eficiência (CLAE) foi eficaz para separação e quantificação adequada das substâncias químicas His, PCA e os isômeros de UCA (trans-UCA e cis-UCA) no estrato córneo dos participantes. O método apresentou-se seletivo e ausente de interferentes, ademais, possuiu linearidade e limites de detecção e quantificação compatíveis com os objetivos dessa investigação. No fototipo I, os níveis de His foram menores em comparação aos demais grupos. Ademais, os níveis dessa mesma substância não apresentaram diferença entre as faixas etárias. Em função da irradiação das amostras, o montante de His aumentou em todos os fototipos. Os níveis de PCA apresentaram-se menores após a irradiação em todos os fototipos de pele. Ainda, as concentrações do PCA foram mais elevadas na faixa etária de 46 a 55 anos de idade. Os níveis de concentração do isômero cis-UCA foram maiores nos participantes com fototipo III, após a irradiação UV. Os níveis de concentração do isômero trans-UCA diminuíram após a irradiação, de forma proporcional à formação de cis-UCA em todos os fototipos. A faixa etária de 46-55 anos de idade obteve níveis significativamente menores de trans-UCA e cis-UCA
Cutaneous hydration occurs, in part, by the components of the Natural Hydration Factor (MFN), originating from the degradation of filagrina, some examples being pyrrolidone-carboxylic acid (PCA), urocanic acid (UCA) and histidine (His). These are present in the stratum corneum (SC). The objective of this research project was to determine, by high efficiency liquid chromatography (HPLC), the PCA, UCA and His in the stratum corneum of participants obtained by tape stripping due to the phototype and age of the research participant. Participants were selected according to age above 18 years, both genders and skin phototype between I and VI, according to Fitzpatrick's classification. The SC samples were obtained from the volar forearm by tape stripping and artificially irradiated. High efficiency liquid chromatography (HPLC) was effective for the separation and proper quantification of the chemicals His, PCA and UCA isomers (trans-UCA and cis-UCA) in the stratum corneum of the participants. The method was selective and absent from interfering, in addition, it had linearity and limits of detection and quantification compatible with the objectives of this investigation. In phototype I, His levels were lower compared to the other groups. Moreover, the levels of this same substance showed no difference between age groups. Due to the irradiation of the samples, the amount of His increased in all phototypes. PCA levels were lower after irradiation in all skin phototypes. Furthermore, PCA concentrations were higher in the age group from 46 to 55 years of age. The concentration levels of the cis-UCA isomer were higher in participants with phototype III after UV irradiation. The concentration levels of the trans-UCA isomer decreased after irradiation, proportionally to the formation of cis-UCA in all phototypes. The age group 46-55 years of age obtained significantly lower levels of trans-UCA and cis-UCA
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Humanos , Masculino , Femenino , Adulto , Piel/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Diagnóstico , Proteínas Filagrina , Anafilaxis Cutánea Pasiva/efectos de los fármacos , Clasificación , Compuestos Químicos , Fluidoterapia/clasificaciónRESUMEN
Three-dimensional skin models, also named 3D skin models, human skin equivalents (HSEs), or Human Epidermal Equivalents (HEEs), have been increasingly used for chemical assessments in terms of efficacy and safety. Considering this, we developed an HEE model using immortalized HaCaT cells, aiming to overcome the limitation of primary tissue source. Our 3D model (HaCaT-HEE) exhibited important markers of cell differentiation (CK10, CK14, involucrin, and filaggrin), although the stratum corneum was shown to be modest. Besides, the model showed a good prediction potential considering membrane permeability, sensitivity, specificity, and accuracy in distinguishing irritant and corrosive effects after exposure to selected chemicals recommended by the OECD protocols. We also validated the formazan determination for the MTT method using High-Performance Liquid Chromatography (HPLC). For that, we considered carry over, linearity, reproducibility/robustness, accuracy, precision, selectivity, and matrix effect, according to the Food and Drug Administration (FDA) guideline. Based on our results, we can conclude that our model has an acceptable predictive value for the safety evaluation of compounds after skin exposure, with the great advantage of being constructed using immortalized cells.
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Cáusticos/toxicidad , Irritantes/toxicidad , Queratinocitos/efectos de los fármacos , Pruebas de Irritación de la Piel/métodos , Alternativas a las Pruebas en Animales , Línea Celular , Permeabilidad de la Membrana Celular/efectos de los fármacos , Epidermis , Proteínas Filagrina , HumanosRESUMEN
Knowing the damage that particulate matter (PM) can cause in skin is important for tightly controlling the release of air pollutants and preventing more serious diseases. This study investigates if such alterations are present in reconstructed human epidermis exposed to coarse air PM. Exposure of reconstructed human epidermis to increasing concentrations (2.2, 8.9, and 17.9 µg/cm2) of standard urban PM over time led to decreased cell viability at 48 hours. The barrier function was shown to be compromised by 24 hours of exposure to high doses (17.9 µg/cm2). Morphological alterations included cytoplasm vacuolization and partial loss of epidermal stratification. Cytokeratin 10, involucrin, loricrin, and filaggrin protein levels were significantly decreased. We confirmed an inflammatory process by IL-1α release and found a significant increase in AQP3 expression. We also demonstrated changes in NOTCH1 and AhR expression of epidermis treated with coarse air PM. The use of hydrogen peroxide altered AQP3 and NOTCH1 expression, and the use of N-acetyl-L-cysteine altered NOTCH1 expression, suggesting that this is a redox-dependent process. These results demonstrate that coarse air PM induces dose-dependent inflammatory response and alterations in protein markers of differentiation and water transport in the epidermis that could ultimately compromise the structural integrity of the skin, promoting or exacerbating various skin diseases.
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Contaminantes Atmosféricos/toxicidad , Epidermis/efectos de los fármacos , Material Particulado/toxicidad , Pérdida Insensible de Agua/efectos de los fármacos , Biomarcadores/análisis , Biomarcadores/metabolismo , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/inmunología , Células Cultivadas , Epidermis/inmunología , Epidermis/metabolismo , Proteínas Filagrina , Humanos , Queratinocitos , Cultivo Primario de Células , Enfermedades de la Piel/inducido químicamente , Enfermedades de la Piel/inmunología , Enfermedades de la Piel/prevención & control , Pérdida Insensible de Agua/inmunologíaRESUMEN
BACKGROUND: Polymorphisms of the filaggrin 2 gene (rs 12568784 and rs 16899374) are associated with persistent atopic dermatitis in African American patients. Filaggrin 2 is a protein with a function similar to filaggrin and also encoded in the epidermal differentiation complex on chromosome 1q21. OBJECTIVE: To evaluate the polymorphisms in the filaggrin 2 gene (rs 12568784 and rs 16899374) in children and adults with atopic dermatitis and to verify the association of these with the severity of the clinical picture, presence of other allergic diseases, and socio-demographic factors. METHOD: The study was carried out with patients and control group. Questionnaires were used to evaluate ethnicity, sex, age, family history, scoring, atopic dermatitis (SCORAD), among other parameters. Genotyping of the filaggrin 2 gene was performed by real-time polymerase chain reaction. RESULTS: Forty-eight patients and 83 controls were evaluated. No correlation was found between the variables studied in patients with atopic dermatitis and polymorphisms, no significant difference between the prevalence of polymorphisms in the patients and in the control group p>0.05. STUDY LIMITS: The exclusive use of self-reported ethnicity information and the sample size. RESULTS: The results of this work can be an incentive for the study of the polymorphisms in atopic dermaititis, considering the characteristic of the Brazilian multi ethnic population. CONCLUSION: This is an unpublished work in Brazil and the first study in the world to have a control group to evaluate alterations in the gene of filaggrin 2.
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Dermatitis Atópica/genética , Polimorfismo Genético/genética , Proteínas S100/genética , Adolescente , Adulto , Brasil , Estudios de Casos y Controles , Niño , Preescolar , Estudios Transversales , Dermatitis Atópica/etnología , Dermatitis Atópica/patología , Femenino , Proteínas Filagrina , Humanos , Lactante , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Índice de Severidad de la Enfermedad , Factores Sexuales , Factores Socioeconómicos , Encuestas y Cuestionarios , Adulto JovenRESUMEN
BACKGROUND: Hydration is an important factor to promote skin barrier function, metabolism, and appearance. In this process, the presence of aquaglyceroporins, envelope and lipid synthesis, and metabolism proteins are essential to provide greater corneocyte cohesion and to form a barrier avoiding transepidermal water loss. OBJECTIVE: We evaluated the effects of a new topical pigment-free agent containing an Anadenanthera colubrina polysaccharide-rich dermocosmetic preparation (ACP) on the aquaporin-3 (AQP-3), filaggrin (FLG), involucrin (INV), glucocerebrosidase (GBA), and elongation of very-long-chain fatty acid (ELOVL) proteins production in skin human fragments, as well as on the transepidermal water loss in a double-blind placebo-controlled clinical trial. METHODS: AQP3, FLG, INV, GBA, and ELOVL3 levels were measured by immunofluorescence analysis in human skin explants. Clinical trial was conducted to evaluate the effects of ACP 1% and ACP 3% on the transepidermal water loss (TEWL). RESULTS: Image and statistical analysis showed that ACP 3% significantly increased at 90% the expression of AQP3. Similarly, ACP 3% was able to promote a significant increase of 68% and 51% in FLG and INV, respectively. ACP 3% produced no effects on the GBA and ELOVL3 proteins. Transepidermal water loss was significantly reduced in human volunteers under treatment with ACP 1% and ACP 3%. CONCLUSION: ACP reduced transepidermal water loss in a clinical trial, promoting human skin hydration. These effects were related to modulation of the AQP3, FLG, and INV as evidenced by immunofluorescence assay. This way, A colubrina polysaccharide-rich phytopharmaceutical preparation is an effective additive product to skin hydration.
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Colubrina , Proteínas Filagrina , Humanos , Preparaciones de Plantas , Polisacáridos/metabolismo , Piel/metabolismo , Agua/metabolismo , Pérdida Insensible de AguaRESUMEN
Aged skin, featured with dryness and wrinkles, has received mounting attention due to its adverse influences on beauty. ß-Sitosterol and vermicularin are two common active ingredients of Thamnolia vermicularis (Sw.) Ach., a traditional Chinese medicine, of which the anti-aging effect has been discovered. Their protective performance against skin aging was assayed by co-culturing with skin cells in this work. Results showed that ß-sitosterol promoted the biosynthesis of hyaluronic acid by increasing the expression of hyaluronic acid synthases in fibroblasts and enhanced the expression of skin barrier functional proteins including aquaporin 3, loricrin, filaggrin and involucrin in keratinocytes, which conduced to the moisture retention within skin. Moreover, vermicularin might function as an anti-wrinkle agent by preventing the loss of collagen type I. Specifically, vermicularin reduced the amount of reactive oxygen species within hydrogen-peroxide-induced fibroblasts; together with suppressing the activation of mitogen-activated protein kinases, it could inhibit the production of matrix metalloproteinases-1. The present research will contribute to the development of the compounds as anti-aging ingredients for future applications in cosmetic formulations and functional food as well as promote further studies of raw materials containing alike compounds.
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Fibroblastos/efectos de los fármacos , Queratinocitos/efectos de los fármacos , Extractos Vegetales/farmacología , Sitoesteroles/farmacología , Envejecimiento de la Piel/efectos de los fármacos , Línea Celular , Proteínas Filagrina , Humanos , Sustancias Protectoras/farmacologíaRESUMEN
PURPOSE: The underlying genetic causes of keratoconus are essentially unknown. Here, we conducted whole-exome sequencing in 2 Brazilian families with keratoconus. METHODS: Whole-exome sequencing was performed on 6 keratoconus-affected individuals of 2 unrelated pedigrees from Southern Brazil. Pathogenic variants were identified in a modified Trio analysis (1 parent and 2 children) using candidate gene filtering. All the affected subjects underwent detailed corneal tomographic evaluation. Clinically relevant variants that were present in affected individuals at minor allele frequencies <1% were examined in the 1000 Genomes Project single nucleotide polymorphism ABraOM and transcription gene (RefSeq and Ensembl) databases. RESULTS: In family 1, a sequence variant in chromosome 1 (q21.3) was observed within the filaggrin gene. All the tested family members shared a heterozygous missense pathogenic variant in the c.4678C>T position. In family 2, exome analysis demonstrated a sequence variant in chromosome 16 (q24.2) within the gene encoding zinc finger protein 469 (ZNF469). Members of family 2 shared a heterozygous missense variant in the c.1489G>A position. In addition, the exomes of the 2 families were examined for shared genetic variants among all affected individuals. Filtering criteria did not identify any rare sequence variants in a single gene segregated in both families. CONCLUSION: Our findings show that a complete genotype-phenotype correlation could not be identified, suggesting that keratoconus is a genetically heterogeneous disease. In addition, we believe that whole-exome sequencing-based segregation analysis is probably not the best strategy for identifying variants in families with isolated keratoconus.
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Secuenciación del Exoma/métodos , Queratocono/genética , Adulto , Córnea/diagnóstico por imagen , Córnea/patología , Femenino , Proteínas Filagrina , Variación Genética/genética , Genómica , Humanos , Masculino , Persona de Mediana Edad , Linaje , Valores de Referencia , Tomografía/métodos , Adulto JovenRESUMEN
Introduction: NLRP3 inflammasome plays a key role in dendritic cells (DC) activation in response to vaccine adjuvants, however we previously showed that it is not properly activated in DC from HIV-infected patients (HIV-DC), explaining, at least in part, the poor response to immunization of these patients. Taking in account that several cytoplasmic receptors are able to activate inflammasome, and that bacterial components are considered as a novel and efficient adjuvant, we postulated that bacterial flagellin (FLG), a natural ligand of NAIP/NLRC4 inflammasome, could rescue the activation of the complex in HIV-DC. Objective: Demonstrate that FLG is able to activate monocyte-derived dendritic cells from HIV-infected individuals better than LPS, and to what extent the entity of inflammasome activation differs between DC from HIV-infected patients and healthy donors. Methods: Monocyte-derived dendritic cells from HIV-infected patients (HIV-DC) and healthy donors (HD-DC) were stimulated with FLG, and inflammasome as well as DC activation (phenotypic profile, cytokine production, autologous lymphocytes activation) were compared. Chemical and genetic inhibitors were used to depict the relative contribution of NLRC4 and NLRP3 in HIV/HD-DC response to FLG. Results: FLG properly activates HD-DC and HIV-DC. FLG induces higher inflammasome activation than LPS in HIV-DC. FLG acts through NLRC4 and NLRP3 in HD-DC, but at a lesser extent in HIV-DC due to intrinsic NLRP3 defect. Conclusions: FLG by-passes NLRP3 defect in HIV-DC, through the activation of NAIP/NLRC4 inflammasome, indicating possible future use of the bacterial component as an efficient adjuvant in immunocompromised individuals.
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Adyuvantes Inmunológicos/farmacología , Proteínas Adaptadoras de Señalización CARD/inmunología , Proteínas de Unión al Calcio/inmunología , Células Dendríticas/inmunología , Flagelina/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Huésped Inmunocomprometido/inmunología , Inflamasomas/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Adulto , Células Dendríticas/patología , Femenino , Proteínas Filagrina , Infecciones por VIH/patología , Humanos , Masculino , Persona de Mediana EdadAsunto(s)
Anemia de Células Falciformes/genética , Predisposición Genética a la Enfermedad/genética , Variación Genética/genética , Úlcera de la Pierna/genética , Proteínas S100/genética , Adulto , Brasil , Estudios de Casos y Controles , Estudios de Cohortes , Femenino , Proteínas Filagrina , Genotipo , Humanos , Inflamación/genética , Masculino , Persona de Mediana Edad , Secuenciación del Exoma/métodosRESUMEN
Inflammatory linear verrucous epidermal nevus and linear psoriasis are sometimes hard to differentiate clinically and pathologically. Although immunohistochemical expression of keratin 10 (K10), K16, Ki-67, and involucrin may be useful for differentiating both entities, these results have been reported in only a few cases. We collected data from 8 patients with inflammatory linear verrucous epidermal nevus, 11 with psoriasis vulgaris, and 8 healthy controls and evaluated immunohistochemical expression of Ki-67, K16, involucrin, and filaggrin among them. Ki-67 and K16 overexpression was similar in inflammatory linear verrucous epidermal nevus and psoriasis vulgaris compared with normal skin. Although staining for involucrin showed discontinuous expression in parakeratotic regions in 4 inflammatory linear verrucous epidermal nevus cases, it was continuous in the other 4 cases and in all psoriasis vulgaris cases. Filaggrin expression was present in hyperkeratotic regions but scarce in parakeratotic areas in both inflammatory linear verrucous epidermal nevus and psoriasis vulgaris. The immunostaining pattern of Ki-67, K16, involucrin, and filaggrin may be insufficient to discriminate inflammatory linear verrucous epidermal nevus from psoriasis vulgaris.
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Proteínas de Filamentos Intermediarios/análisis , Queratina-16/análisis , Antígeno Ki-67/análisis , Nevo Sebáceo de Jadassohn/diagnóstico , Precursores de Proteínas/análisis , Psoriasis/diagnóstico , Adolescente , Adulto , Anciano , Biomarcadores/análisis , Estudios de Casos y Controles , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Proteínas Filagrina , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Nevo Sebáceo de Jadassohn/patología , Psoriasis/patologíaRESUMEN
The Brazilian AIDS epidemic has been characterized by an increasing rate of BF1 recombinants and so far eight circulating recombinant forms/CRFs_BF1 have been described countrywide. In this study, pol sequences (protease/PR, reverse transcriptase/RT) of 87 BF1 mosaic isolates identified among 828 patients living in six Brazilian States from three geographic regions (Central West, North, Northeast) were analyzed. Phylogenetic and bootscan analyses were performed to investigate the evolutionary relationship and mosaic structure of BF1 isolates. Those analyses showed that 20.7% of mosaics (18 out of 87) were CRFs-like isolates, mostly represented by CRF28/CRF29_BF-like viruses (14 out of 18). We also identified five highly supported clusters that together comprise 42 out of 87 (48.3%) BF1 sequences, each cluster containing at least five sequences sharing a similar mosaic structure, suggesting possible new unidentified CRFs_BF1. The divergence time of these five potential new CRFs_BF1 clusters was estimated using a Bayesian approach and indicate that they probably originated between the middle 1980s and the middle 1990s. DNA was extracted from whole blood and four overlapping fragments were amplified by PCR providing full/near full length genomes (FLG/NFLG) and partial genomes. Eleven HIV-1 isolates from Cluster # 5 identified in epidemiologically unlinked individuals living in Central West and North regions provided FLG/NFLG/partial genome sequences with identical mosaic structure. These viruses differ from any known CRF_BF1 reported to date and were named CRF90_BF1 by the Los Alamos National Laboratory. This is the 9th CRF_BF1 described in Brazil and the first one identified in Central West and North regions. Our results highlight the importance of continued molecular screening and surveillance studies, especially of full genome sequences to understand the evolutionary dynamics of the HIV-1 epidemic in a country of continental dimensions as Brazil.
Asunto(s)
Genoma Viral , Infecciones por VIH/virología , VIH-1/genética , Productos del Gen pol del Virus de la Inmunodeficiencia Humana/genética , Adulto , Teorema de Bayes , Brasil/epidemiología , Análisis por Conglomerados , Evolución Molecular , Femenino , Proteínas Filagrina , Genotipo , Infecciones por VIH/epidemiología , VIH-1/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Filogenia , ARN Viral/aislamiento & purificación , ARN Viral/metabolismo , Recombinación Genética , Análisis de Secuencia de ADN , Adulto Joven , Productos del Gen pol del Virus de la Inmunodeficiencia Humana/clasificaciónRESUMEN
Atopic dermatitis is a chronic inflammatory skin disease with a complex pathogenesis, where changes in skin barrier and imbalance of the immune system are relevant factors. The skin forms a mechanic and immune barrier, regulating water loss from the internal to the external environment, and protecting the individual from external aggressions, such as microorganisms, ultraviolet radiation and physical trauma. Main components of the skin barrier are located in the outer layers of the epidermis (such as filaggrin), the proteins that form the tight junction (TJ) and components of the innate immune system. Recent data involving skin barrier reveal new information regarding its structure and its role in the mechanic-immunological defense; atopic dermatitis (AD) is an example of a disease related to dysfunctions associated with this complex.
Asunto(s)
Dermatitis Atópica/inmunología , Epidermis/inmunología , Proteínas de Filamentos Intermediarios/inmunología , Dermatitis Atópica/fisiopatología , Epidermis/fisiopatología , Proteínas Filagrina , Humanos , Inmunidad Innata , Proteínas de Filamentos Intermediarios/análisis , Receptores de Reconocimiento de Patrones/análisis , Receptores de Reconocimiento de Patrones/inmunología , Uniones Estrechas/inmunologíaRESUMEN
The prevalence rates of anti-citrullinated protein/peptide antibodies (ACPAs) were investigated in a cohort of juvenile idiopathic arthritis (JIA) patients, and their diagnostic performances were compared. ACPAs, including anti-cyclic citrullinated peptide IgG (anti-CCP), anti-CCP IgG/IgA (anti-CCP3.1), citrullinated recombinant rat filaggrin antibodies (CPA), anti-mutated citrullinated vimentin (anti-MCV), and antibodies to citrullinated human IgG-derived peptides (RA/CP), were measured in the sera from 81 JIA patients. Serum samples from 55 children with other joint diseases or viral infections and 49 healthy donors were tested as controls. Of the 81 JIA patients, 7 (8.6%), 8 (9.9%), 17 (21.0%), 23 (28.4%), and 18 (22.2%) were found to be positive for anti-CCP, anti-CCP3.1, CPA, anti-MCV, and RA/CP, respectively, with specificities of 98.1, 95.1, 93.3, 84.6, and 86.5%. Analysis by subtype revealed that 7/7 (100%) of RF-positive polyarticular JIA patients tested positive at high serum levels for anti-MCV or RA/CP, and 5/7 (71.4%) were positive for anti-CCP, anti- CCP3.1, or CPA (P < 0.001, compared with controls). Eighteen of 81 JIA patients demonstrated joint erosions on radiographs and erosive arthritis occurred more often in ACPAs positive patients (P < 0.01). Our findings indicate that although ACPAs are not satisfactory screening biomarkers for JIA due to low sensitivity, ACPA measurement can aid in diagnosing RF-positive polyarticular JIA and identifying JIA patients with severe bone involvement. The diagnostic performance of each ACPA in JIA is different, and the careful selection of assays is necessary.