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1.
J Phys Chem B ; 128(1): 45-55, 2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38154791

RESUMO

Environmental pollution by man-made toxic and persistent organic compounds, found throughout the world in surface and groundwater, has various negative effects on aquatic life systems and even humans. Therefore, it is important to develop and improve water treatment technologies capable of removing such substances from wastewater and purifying drinking water. The two substances investigated are the widely used painkiller diclofenac and a member of the class of "forever chemicals", perfluorobutanesulfonate. Both are known to have serious negative effects on living organisms, especially under long-term exposure, and are detectable in human hair, suggesting adsorption to a part of the hair fiber complex. In this study, a human hair keratin dimer is investigated for its ability to absorb diclofenac and perfluorobutanesulfonate. Initial predictions for binding sites are obtained via molecular docking and subjected to molecular dynamics simulations for more than 1 µs. The binding affinities obtained by the linear interaction energy method are high enough to motivate further research on human hair keratins as a sustainable, low-cost, and easily allocatable filtration material.


Assuntos
Diclofenaco , Fluorocarbonos , Queratinas Específicas do Cabelo , Ácidos Sulfônicos , Humanos , Queratinas Específicas do Cabelo/análise , Adsorção , Simulação de Acoplamento Molecular , Cabelo/química
2.
Cells ; 12(23)2023 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-38067138

RESUMO

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced, and 599 candidate genes exhibited differential expression between DSG3-overexpressing and matched control lines, with 12 genes highly significantly differentially expressed, including 9 upregulated and 3 downregulated. Genes with known implications in cancer, such as MMP-13, KRT84, OLFM4, GJA1, AMOT and ADAMTS1, were strongly linked to DSG3 overexpression. Gene ontology analysis indicated that the DSG3-associated candidate gene products participate in crucial cellular processes such as junction assembly, focal adhesion, extracellular matrix formation, intermediate filament organisation and keratinocyte differentiation. Validation of RNA-Seq was performed through RT-qPCR, Western blotting and immunofluorescence analyses. Furthermore, using transmission electron microscopy, we meticulously examined desmosome morphology and revealed a slightly immature desmosome structure in DSG3-overexpressing cells compared to controls. No changes in desmosome frequency and diameter were observed between the two conditions. This study underscores intricate and multifaceted alterations associated with DSG3 in oral squamous carcinoma cells, implying a potential oncogenic role of this gene in biological processes that enable cell communication, motility and survival.


Assuntos
Carcinoma de Células Escamosas , Neoplasias Bucais , Humanos , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Desmogleína 3/genética , Desmogleína 3/análise , Desmogleína 3/metabolismo , Desmossomos/metabolismo , Perfilação da Expressão Gênica , Queratinócitos/metabolismo , Queratinas Específicas do Cabelo/análise , Queratinas Específicas do Cabelo/genética , Queratinas Específicas do Cabelo/metabolismo , Queratinas Tipo II/análise , Queratinas Tipo II/genética , Queratinas Tipo II/metabolismo , Neoplasias Bucais/genética , Neoplasias Bucais/metabolismo , Oncogenes , Transcriptoma
3.
Acta Biomater ; 168: 440-457, 2023 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-37479159

RESUMO

Cosmetics for perming hair are commonly used but have negative impacts on hair fibers. Repairing damaged hair with conditioners, hair oil, and hair masks can provide relief but cannot prevent injuries. Recent research has shown that proteins and amino acids can remodel hair's disulfide bonds. However, the permeation ability of proteins is limited, and amino acids may disrupt the secondary structure of hair keratins. Our study demonstrates that peptides can be safely, efficiently, and promisingly used for hair perming. A bioinspired peptide, PepACS (PepA-PepC-SPB), was designed through bioinformatics. It can interact with keratin's sulfhydryl group in situ to remodel disulfide bonds without affecting hair fiber's tensile properties. The potential of PepACS to repair cuticle injuries is also observed through scanning electron microscope visualization. Besides, linking PepACS with mCherry enables hair dyeing. This research suggests that biomaterials can be applied in the hair care industry. STATEMENT OF SIGNIFICANCE: Chemical perming products can have negative impacts on people's health and hair fibers, making it essential to explore alternative methods. Peptides treatment is a promising option, but synthesizing sulfur-rich short peptides for hair perming has not been demonstrated before. In this paper, we utilized bioinformatics to design bio-inspired peptides that can interact with hair keratins and form curled shapes. Our study demonstrates that bioinformatics tools can be utilized to design bioinspired peptides with unique functions. Sulfur-rich short peptides can be heterologously expressed with fusion strategies, and PepACS can securely bind hair fibers through disulfide bonds. Importantly, perming hair with 0.01% PepACS maintains the mechanical properties of hair, and dyeing hair with the fusion protein PepACS_mCh can be facilitated by ethanol. These findings suggest that the strategy of perming and dyeing hair through peptides is non-injurious, and the peptides used for repairing hair damage show tremendous potential.


Assuntos
Tinturas para Cabelo , Queratinas Específicas do Cabelo , Humanos , Queratinas Específicas do Cabelo/análise , Queratinas Específicas do Cabelo/metabolismo , Tinturas para Cabelo/análise , Tinturas para Cabelo/química , Tinturas para Cabelo/metabolismo , Proteínas/metabolismo , Peptídeos/metabolismo , Aminoácidos/análise , Cabelo/química , Dissulfetos/metabolismo
4.
Int J Cosmet Sci ; 44(5): 588-601, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35916243

RESUMO

OBJECTIVE: The objective of our research was to investigate the heat-protecting effect of a product ex vivo and in vivo on human hair fibres. METHODS: A preparatory study was carried out in order to determine an optimal threshold of thermal stress. For this, the structure of cross-sections of the hair fibre was observed by optical microscopy. Then, Scanning Electron Microscopy (SEM) and Confocal Raman Spectroscopy (CRS) were applied to analyse ex vivo and in vivo morphological and molecular damage in hair structure after heat stress. Finally, in vivo tests were used to collect consumer perception. RESULTS: The preparatory study enabled us to determine an optimal stress threshold of 10 heating cycle for SEM and 5 heating cycle for CRS. Based on spectral hierarchical classification using Ward's clustering algorithm, the ex vivo Raman results show that the spectral signature of the hair treated and heated is very close to the negative control. This shows that the product preserves the keratin structure after thermal stress. These results were also confirmed by an in vivo Raman analysis performed on hair samples from 5 donors. In concordance with Raman results, SEM shows that treated hair presents lesser "bubbles" and "crackling" on the hair surface. Finally, the in vivo studies proved that hair was more protected from heat. CONCLUSION: The authors concluded that the product shows protective properties with respect to morphological and molecular heat damage. We also demonstrate that the product promotes the α-helix keratin conformation and preserves the S-S disulfide bands.


OBJECTIF: L'objectif de notre étude est d'évaluer ex vivo et in vivo l'effet thermoprotecteur d'un produit sur les fibres capillaires humaines. MÉTHODES: Une étude préparatoire a été réalisée afin de déterminer un seuil optimal du stress thermique. Pour cela, la structure des coupes transversales des cheveux a été observée par microscopie optique. Ensuite, la microscopie électronique à balayage (MEB) et la spectroscopie confocale Raman (SCR) ont été appliquées pour analyser les dommages morphologiques et moléculaires (ex vivo et in vivo) de la structure du cheveu après un stress thermique. Enfin, des tests in vivo ont été réalisés pour recueillir la perception des consommateurs. RÉSULTATS: L'étude préparatoire nous a permis de déterminer un seuil de stress thermique optimal correspondant à 10 cycles de chauffage pour la MEB et 5 cycles de chauffage pour la SCR. Basés sur une classification hiérarchique utilisant l'algorithme de Ward, les résultats Raman « ex vivo ¼ montrent que la signature spectrale des cheveux traités et chauffés est très proche du témoin négatif. Cela montre que le produit préserve la structure de la kératine après un stress thermique. Ces résultats ont également été confirmés par une analyse Raman « in vivo ¼ réalisée sur des échantillons de cheveux de 5 donneurs. En concordance avec les résultats Raman, la MEB montre que les cheveux traités présentent moins de « bulles ¼ et de « craquelures ¼ à la surface des cheveux. Enfin, l'étude in vivo a prouvé que les cheveux sont mieux protégés de la chaleur. CONCLUSION: Les auteurs ont conclu que le produit présente des propriétés protectrices vis-à-vis des dommages thermiques morphologiques et moléculaires. Nous avons démontré également que le produit favorise la conformation de la kératine en hélice-α et préserve les bandes disulfures S-S.


Assuntos
Preparações para Cabelo , Queratinas Específicas do Cabelo , Cabelo/química , Humanos , Queratinas Específicas do Cabelo/análise , Microscopia Eletrônica de Varredura , Análise Espectral Raman/métodos
5.
Curr Pharm Des ; 28(10): 841-851, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35034588

RESUMO

BACKGROUND: Keratin is among the most abundant structural proteins of animal origin, however it remains broadly underutilized. OBJECTIVE: Bioinformatic investigation was performed to evaluate selected keratins originating from mass-produced waste products, i.e., chicken feathers and pig hair, as potential sources of bioactive peptides. METHODS: Pepsin, trypsin, chymotrypsin, papain, and subtilisin were used for in silico keratinolysis with the use of "Enzyme(s) action" and fragmentomic analysis of theoretical products was performed using "Profiles of potential biological activity" in BIOPEP-UWM database of bioactive peptides. Bioactivity probability calculation and toxicity prediction of the peptides obtained were estimated using PeptideRanker and ToxinPred tools, respectively. RESULTS: Our results showed that the keratins are a potential source of a variety of biopeptides, including dipeptidyl peptidase IV, angiotensin converting enzyme, prolyl endopeptidase inhibitory and antioxidative. Papain and subtilisin were found to be the most appropriate enzymes for keratin hydrolysis. This study presents possible structures of keratin-derived bioactive peptides that have not been previously described. CONCLUSION: Our data suggest additional in vitro and in vivo studies to verify theoretical predictions and further investigate the possibility of using keratin-rich waste as a source of peptide nutraceuticals.


Assuntos
Plumas , Queratinas Específicas do Cabelo , Animais , Galinhas , Plumas/química , Queratinas Específicas do Cabelo/análise , Papaína/análise , Peptídeos/química , Subtilisinas/metabolismo , Suínos
6.
J Biomed Mater Res A ; 110(1): 92-104, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34254735

RESUMO

Human hair keratin (HHK) has been successfully explored as raw materials for three-dimensional scaffolds for soft tissue regeneration due to its excellent biocompatibility and bioactivity. However, none of the reported HHK based scaffolds is able to replicate the strain-stiffening capacity of living tissues when responding to large deformations. In the present study, strain-stiffening property was achieved in scaffolds fabricated from HHK via a synergistic effect of well-defined, aligned microstructure and chemical crosslinking. Directed ice-templating method was used to fabricate HHK-based scaffolds with highly aligned (anisotropic) microstructure while oxidized dopamine (ODA) was used to crosslink covalently to HHKs. The resultant HHK-ODA scaffolds exhibited strain-stiffening behavior characterized by the increased gradient of the stress-strain curve after the yield point. Both ultimate tensile strength and the elongation at break were enhanced significantly (~700 kPa, ~170%) in comparison to that of HHK scaffolds lacking of aligned microstructure or ODA crosslinking. In vitro cell culture studies indicated that HHK-ODA scaffolds successfully supported human dermal fibroblasts (HDFs) adhesion, spreading and proliferation. Moreover, anisotropic HHK-ODA scaffolds guided cell growth in alignment with the defined microstructure as shown by the highly organized cytoskeletal networks and nuclei distribution. The findings suggest that HHK-ODA scaffolds, with strain-stiffening properties, biocompatibility and bioactivity, have the potential to be applied as biomimetic matrices for soft tissue regeneration.


Assuntos
Dopamina , Queratinas Específicas do Cabelo , Anisotropia , Cabelo/química , Humanos , Queratinas Específicas do Cabelo/análise , Queratinas Específicas do Cabelo/química , Resistência à Tração , Engenharia Tecidual/métodos , Tecidos Suporte/química
7.
Virchows Arch ; 479(3): 515-521, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34218288

RESUMO

Urothelial carcinoma is subdivided into luminal (L), basal (B), and p53-wild-type (WT) molecular subtypes, with basal and p53-WT groups showing more aggressive course and poor treatment response, respectively. The literature on molecular subtypes of UC includes a mixture of different stages. We investigated the molecular profile and outcome of pure cohort of muscle invasive bladder carcinoma (MIBC) considering two distinct patterns of muscularis propria (MP) invasion. Forty-three cystectomies harboring stage pT2 were retrospectively identified in 18 years. MP invasion was subclassified into patterns 1 (tumor encasing intact detrusor muscle bundles) and 2 (tumor dissecting/replacing detrusor muscle). Using IHC, B/L phenotypes, p53, and Ki67 were assessed, and survival data was collected. Pattern 1 invasion was noted in 16 (37%) and pattern 2 in 27 (63%), with mean age of pattern 1 being 10 years younger. B/L phenotypes were successfully determined in 83.7%; 48.8% and 34.8% revealed L and B phenotypes, respectively (indeterminate phenotype in 16.4%). Pattern 1 was associated with L phenotype (GATA3 and HER-2 expressions: p = 0.02 & p = 0.04, respectively). Ki67 ≥ 5/10HPF was noted in pattern 2 and B phenotype (p = 0.03). B phenotype showed association with p53-WT (p = 0.007). In median follow-up of 60.7 months, 63.6% of pattern 1 cases were alive without disease compared to 32% of pattern 2 (not significant). A panel of CK20 and GATA3 for luminal and CK5/6 and CK14 for basal subtypes can provide reliable molecular classification in UC. Also, morphology of MIBC can predict the molecular phenotype and the behavior of the UC.


Assuntos
Biomarcadores Tumorais/análise , Carcinoma/química , Neoplasias da Bexiga Urinária/química , Urotélio/química , Idoso , Carcinoma/classificação , Carcinoma/patologia , Carcinoma/cirurgia , Cistectomia , Bases de Dados Factuais , Feminino , Fator de Transcrição GATA3/análise , Humanos , Imuno-Histoquímica , Queratina-14/análise , Queratina-20/análise , Queratina-5/análise , Queratinas Específicas do Cabelo/análise , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Fenótipo , Valor Preditivo dos Testes , Estudos Retrospectivos , Resultado do Tratamento , Neoplasias da Bexiga Urinária/classificação , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/cirurgia , Urotélio/patologia , Urotélio/cirurgia
8.
ACS Appl Bio Mater ; 4(8): 6195-6202, 2021 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35006898

RESUMO

The coloration of human hair keratin fibers has long involved the oxidative coupling of primarily aromatic amines and phenols inside the fibers with the aid of harsh agents such as H2O2 and NH4OH. Further, the traditional process has exposed millions of consumers and their hairstylists to toxic substances such as skin sensitizers. While alternative hair dyeing processes have been explored, they fail to be competitive with the traditional method, for reasons including impracticality and limited colors achievable. In the present study, we developed an approach to imparting color to human hair fibers that involves entrapping colorants inside hair fibers by forming chelated monoazo dyes in situ. Dyes employed were based on monoarylide, arylazopyrazolone, and arylazonaphthol families, which display yellow, orange, and magenta colors on dyed hair. The dyes were applied at 40 °C without the use of oxidants and alkali associated with current commercial hair dyes, with the best dye uptake observed when the arylazonaphthol dye was employed. The dyed hair fibers showed good durability to washing, and treatment of these fibers with Al3+ or Fe3+ ions at 40 °C led to the rapid in situ formation of 1:2 metal/dye structures. In addition, the dyed hair was soft, indicating that chelated dye occupies the interior of the fibers rather than the surface. Such an approach can be applied to the coloration of other materials, including textiles.


Assuntos
Tinturas para Cabelo , Queratinas Específicas do Cabelo , Proteínas do Citoesqueleto/análise , Cabelo/química , Tinturas para Cabelo/análise , Humanos , Peróxido de Hidrogênio/análise , Íons/análise , Queratinas Específicas do Cabelo/análise , Metais/análise
9.
An. bras. dermatol ; 95(3): 278-282, May-June 2020. tab, graf
Artigo em Inglês | LILACS, Coleciona SUS | ID: biblio-1130884

RESUMO

Abstract Background: The hair follicle is a unique structure, one of the most dynamic structures in mammalians, which can reproduce in every new cycle all the mechanism involved in its fetal development. Although a lot of research has been made about the human hair follicle much less has been discovered about the importance of the cytokeratins (CKs) in its development. Objective: Study the immunohistochemical pattern of epithelial CKs during human hair follicle development. Methods: We performed an immunohistochemical study using fresh post-mortem skin biopsies of human fetuses between 4 and 25 weeks of gestational age to study the expression of cytokeratins (CKs): CK1, CK10, CK13, CK14, CK16 and CK20 during human hair follicle fetal development. Study limitations: Restrospective study with a good number of makers but with a small population. Results/conclusion: We found that, the CKs were expressed in an intermediate time during follicular development. The epithelial CKs (CK1, CK14, CK10, CK13) and the epithelial CKs with a proliferative character such as CK16 were expressed first, as markers of cellular maturation and follicular keratinization. At a later phase, CK20 was expressed in more developed primitive hair follicles as previously discussed in literature.


Assuntos
Folículo Piloso/cirurgia , Folículo Piloso/crescimento & desenvolvimento , Queratinas Específicas do Cabelo/análise , Imuno-Histoquímica , Estudos Retrospectivos , Fatores Etários , Idade Gestacional , Desenvolvimento Fetal , Anticorpos Monoclonais/análise
10.
An Bras Dermatol ; 95(3): 278-282, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32299738

RESUMO

BACKGROUND: The hair follicle is a unique structure, one of the most dynamic structures in mammalians, which can reproduce in every new cycle all the mechanism involved in its fetal development. Although a lot of research has been made about the human hair follicle much less has been discovered about the importance of the cytokeratins (CKs) in its development. OBJECTIVE: Study the immunohistochemical pattern of epithelial CKs during human hair follicle development. METHODS: We performed an immunohistochemical study using fresh post-mortem skin biopsies of human fetuses between 4 and 25 weeks of gestational age to study the expression of cytokeratins (CKs): CK1, CK10, CK13, CK14, CK16 and CK20 during human hair follicle fetal development. STUDY LIMITATIONS: Restrospective study with a good number of makers but with a small population. RESULTS/CONCLUSION: We found that, the CKs were expressed in an intermediate time during follicular development. The epithelial CKs (CK1, CK14, CK10, CK13) and the epithelial CKs with a proliferative character such as CK16 were expressed first, as markers of cellular maturation and follicular keratinization. At a later phase, CK20 was expressed in more developed primitive hair follicles as previously discussed in literature.


Assuntos
Folículo Piloso/citologia , Folículo Piloso/crescimento & desenvolvimento , Queratinas Específicas do Cabelo/análise , Fatores Etários , Anticorpos Monoclonais/análise , Desenvolvimento Fetal , Idade Gestacional , Humanos , Imuno-Histoquímica , Estudos Retrospectivos
11.
Biosci Rep ; 40(4)2020 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-32181476

RESUMO

AIMS: Oral squamous cell carcinoma (OSCC) is a common oral cancer; however, current therapeutic approaches still show limited efficacy. Our research aims to explore effective biomarkers related to OSCC. MAIN METHODS: Gene expression profiles of paired OSCC tumor and paracancerous samples from The Cancer Genome Atlas (TCGA) were analyzed. mRNA and protein levels of KRT84 in OSCC cell line HSC-3 were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. KRT84 protein levels in OSCC tumor samples of different stages were determined by immunohistochemistry. Overall survival (OS) of OSCC samples was evaluated and association of multiple factors with OS was assessed. KEY FINDINGS: Compared with paracancerous samples, 4642 DEGs were identified in OSCC tumor samples. Among them, KRT84 expression level in OSCC tumor tissues was obviously decreased, which was validated in HSC-3 cells. KRT84 expression level showed decreasing tendency with the increase of tumor grade and stage. Patients with low KRT84 expression level had inferior OS independently of multiple factors. Besides, antigen processing and presentation pathway were significantly activated in OSCC samples with high KRT84 expression. Elevated KRT84 mRNA as well as protein levels were confirmed by RT-qPCR and Western blot in OSCC and normal cell lines, and immunohistochemistry in OSCC tumor and paracancerous tissues. SIGNIFICANCE: Our study suggests KRT84 as a tumor suppressor and good prognostic indicator for OSCC, which might be significant for OSCC diagnosis and treatment.


Assuntos
Biomarcadores Tumorais/metabolismo , Queratinas Específicas do Cabelo/metabolismo , Queratinas Tipo II/metabolismo , Neoplasias Bucais/patologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Proteínas Supressoras de Tumor/metabolismo , Biomarcadores Tumorais/análise , Linhagem Celular Tumoral , Conjuntos de Dados como Assunto , Feminino , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Queratinas Específicas do Cabelo/análise , Queratinas Tipo II/análise , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/patologia , Neoplasias Bucais/diagnóstico , Neoplasias Bucais/mortalidade , Prognóstico , Carcinoma de Células Escamosas de Cabeça e Pescoço/diagnóstico , Carcinoma de Células Escamosas de Cabeça e Pescoço/mortalidade , Análise de Sobrevida , Proteínas Supressoras de Tumor/análise
12.
Nutrients ; 11(12)2019 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-31847069

RESUMO

Hair disorders may considerably impact the social and psychological well-being of an individual. Recent advances in the understanding the biology of hair have encouraged the research and development of novel and safer natural hair growth agents. In this context, we have previously demonstrated-at both preclinical and clinical level-that an Annurca apple-based dietary supplement (AMS), acting as a nutraceutical, is endowed with an intense hair-inductive activity (trichogenicity), at once increasing hair tropism and keratin content. Herein, in the framework of preclinical investigations, new experiments in primary human models of follicular keratinocytes and dermal papilla cells have been performed to give an insight around AMS biological effects on specific hair keratins expression. As well as confirming the biocompatibility and the antioxidant proprieties of our nutraceutical formulation, we have proven an engagement of trichokeratins production underlying its biological effects on human follicular cells. Annurca apples are particularly rich in oligomeric procyanidins, natural polyphenols belonging to the broader class of bioflavonoids believed to exert many beneficial health effects. To our knowledge, none of the current available remedies for hair loss has hitherto shown to stimulate the production of hair keratins so clearly.


Assuntos
Folículo Piloso , Queratinas Específicas do Cabelo , Malus , Extratos Vegetais/farmacologia , Antioxidantes/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Suplementos Nutricionais , Flavonoides , Folículo Piloso/citologia , Folículo Piloso/efeitos dos fármacos , Folículo Piloso/metabolismo , Humanos , Queratinas Específicas do Cabelo/análise , Queratinas Específicas do Cabelo/metabolismo , Modelos Biológicos
13.
Biotechniques ; 64(4): 170-176, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29661011

RESUMO

A simple method for extracting protein from human anagen (i.e., actively growing hair stage) head hairs was developed in this study for cases of limited sample availability and/or studies of specific micro-features within a hair. The distinct feature segments of the hair from one donor were divided lengthwise (i.e., each of ∼200-400 µm) and then pooled for three individual hairs to form a total of eight composite hair samples (i.e., each of ∼1 mm or less in total length). The proteins were extracted, digested using trypsin, and characterized via nano-flow liquid chromatography tandem-mass spectrometry (nLC-MS/MS). A total of 63 proteins were identified from all eight protein samples analyzed of which 60% were keratin and keratin-associated proteins. The major hair keratins identified are consistent with previous studies using fluorescence in situ hybridization and nLC-MS/MS while requiring over 400-8000-fold less sample. The protein extraction method from micro-sized human head hairs described in this study will enable proteomic analysis of biological evidence for cases of limited sample availability and will complement hair research. For example, research seeking to develop alternative non-DNA based techniques for comparing questioned to known hairs, and understanding the biochemistry of hair decomposition.


Assuntos
Cabelo/química , Queratinas Específicas do Cabelo/análise , Proteínas/análise , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos , Adulto , Cromatografia Líquida/métodos , Humanos , Masculino , Proteólise , Tripsina/química
14.
Sci Rep ; 8(1): 1599, 2018 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-29371649

RESUMO

Human hair is laminar-fibrous tissue and an evolutionarily old keratinization product of follicle trichocytes. Studies on the hair proteome can give new insights into hair function and lead to the development of novel biomarkers for hair in health and disease. Human hair proteins were extracted by detergent and detergent-free techniques. We adopted a shotgun proteomics approach, which demonstrated a large extractability and variety of hair proteins after detergent extraction. We found an enrichment of keratin, keratin-associated proteins (KAPs), and intermediate filament proteins, which were part of protein networks associated with response to stress, innate immunity, epidermis development, and the hair cycle. Our analysis also revealed a significant deamidation of keratin type I and II, and KAPs. The hair shafts were found to contain several types of histones, which are well known to exert antimicrobial activity. Analysis of the hair proteome, particularly its composition, protein abundances, deamidated hair proteins, and modification sites, may offer a novel approach to explore potential biomarkers of hair health quality, hair diseases, and aging.


Assuntos
Cabelo/química , Histonas/análise , Queratinas Específicas do Cabelo/análise , Proteoma/análise , Humanos , Processamento de Proteína Pós-Traducional
15.
Hum Pathol ; 46(3): 471-5, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25543160

RESUMO

Hyalinizing clear cell carcinoma (HCCC) has only been described in salivary glands of the head and neck. We report a 38-year-old man with a 2.6-cm lung tumor that was growing in a peribronchial location and had morphologic features of HCCC. The tumor cells expressed cytokeratin 7 and keratin AE1/AE3, and the vast majority of tumor cells marked also with p63 and p40. They were negative for cytokeratin 20, S-100, smooth muscle actin, napsin A, and thyroid transcription factor-1. Fluorescence in situ hybridization revealed Ewing Sarcoma Breakpoint Region 1 (EWSR1) rearrangement, and reverse-transcription polymerase chain reaction confirmed the presence of the EWSR1-Activating Transcription Factor 1 (ATF1) fusion transcript, which was subsequently sequenced. The morphologic, immunophenotypic, cytogenetic, and molecular findings together with the patient's history and location of the tumor support a diagnosis of primary pulmonary HCCC of bronchial submucosal gland origin. It is our understanding that this is the first report of HCCC arising as a primary tumor outside the head and neck region.


Assuntos
Adenocarcinoma de Células Claras/patologia , Carcinoma Broncogênico/patologia , Neoplasias Pulmonares/patologia , Adenocarcinoma de Células Claras/química , Adenocarcinoma de Células Claras/genética , Adulto , Proteínas de Ligação a Calmodulina/química , Proteínas de Ligação a Calmodulina/genética , Carcinoma Broncogênico/química , Carcinoma Broncogênico/genética , Rearranjo Gênico , Humanos , Epitopos Imunodominantes/análise , Imuno-Histoquímica , Queratina-7/análise , Queratinas Específicas do Cabelo/análise , Neoplasias Pulmonares/química , Neoplasias Pulmonares/genética , Masculino , Proteínas de Membrana/análise , Fragmentos de Peptídeos/análise , Proteína EWS de Ligação a RNA , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética
16.
PLoS One ; 9(4): e93607, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24714551

RESUMO

Pure hair and nail ectodermal dysplasia (PHNED) comprises a heterogeneous group of rare heritable disorders characterized by brittle hair, hypotrichosis, onychodystrophy and micronychia. Autosomal recessive (AR) PHNED has previously been associated with mutations in either KRT85 or HOXC13 on chromosome 12p11.1-q14.3. We investigated a consanguineous Pakistani family with AR PHNED linked to the keratin gene cluster on 12p11.1 but without detectable mutations in KRT85 and HOXC13. Whole exome sequencing of affected individuals revealed homozygosity for a rare c.821T>C variant (p.Phe274Ser) in the KRT74 gene that segregates AR PHNED in the family. The transition alters the highly conserved Phe274 residue in the coil 1B domain required for long-range dimerization of keratins, suggesting that the mutation compromises the stability of intermediate filaments. Immunohistochemical (IHC) analyses confirmed a strong keratin-74 expression in the nail matrix, the nail bed and the hyponychium of mouse distal digits, as well as in normal human hair follicles. Furthermore, hair follicles and epidermis of an affected family member stained negative for Keratin-74 suggesting a loss of function mechanism mediated by the Phe274Ser substitution. Our observations show for the first time that homozygosity for a KRT74 missense variant may be associated with AR PHNED. Heterozygous KRT74 mutations have previously been associated with autosomal dominant woolly hair/hypotrichosis simplex (ADWH). Thus, our findings expand the phenotypic spectrum associated with KRT74 mutations and imply that a subtype of AR PHNED is allelic with ADWH.


Assuntos
Fissura Palatina/genética , Displasia Ectodérmica/genética , Cabelo/patologia , Hipotricose/genética , Deficiência Intelectual/genética , Queratinas Específicas do Cabelo/genética , Queratinas Tipo II/genética , Mutação de Sentido Incorreto , Unhas/patologia , Sindactilia/genética , Alelos , Sequência de Aminoácidos , Animais , Fissura Palatina/patologia , Consanguinidade , Displasia Ectodérmica/patologia , Cabelo/metabolismo , Homozigoto , Humanos , Hipotricose/patologia , Deficiência Intelectual/patologia , Queratinas Específicas do Cabelo/análise , Queratinas Tipo II/análise , Camundongos , Dados de Sequência Molecular , Unhas/metabolismo , Linhagem , Sindactilia/patologia
17.
J Cutan Pathol ; 41(5): 427-36, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24673383

RESUMO

BACKGROUND: Pilomatricomas are tumors that emulate the differentiation of matrix cells of the hair follicle, showing cortical differentiation, with sequential expression of K35 and K31 keratins. Beta-catenin gene is frequently mutated in pilomatricoma, leading to beta-catenin nuclear accumulation, and to downstream expression of LEF1. Skin matrical tumors other than pilomatricoma are very rare, and comprise purely matrical tumors and focally matrical tumors. We aimed at studying cortical differentiation, beta-catenin pathway and expression of the follicular stem-cell marker PHLDA1 in a series of matrical tumors other than pilomatricoma. METHODS: In 36 prospectively collected tumors, K31, K35, CK17, LEF1, HOXC13, beta-catenin and PHLDA1 expressions were evaluated. Five pilomatricomas were used as controls. RESULTS: In 18 purely matrical tumors (11 matrical carcinomas, 4 melanocytic matricomas, 3 matricomas) and 18 focally matrical tumors (11 basal cell carcinomas, 3 trichoepithelioma/trichoblastomas, 4 others), sequential K35, HOXC13 and K31 expressions were found, indicating cortical differentiation. Germinative matrix cells were always CK17-, and showed nuclear beta-catenin accumulation, with LEF1 and PHLDA1 expressions. CONCLUSIONS: Nuclear beta-catenin and LEF1 expression was highly conserved in matrical tumors, and suggested a common tumorigenesis driven by Wnt pathway activation. PHLDA1 was consistently expressed in matrical tumors and in areas of matrical differentiation.


Assuntos
Doenças do Cabelo/patologia , Queratinas Específicas do Cabelo/biossíntese , Neoplasias Cutâneas/patologia , Fatores de Transcrição/biossíntese , beta Catenina/biossíntese , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/análise , Feminino , Doenças do Cabelo/metabolismo , Humanos , Imuno-Histoquímica , Queratinas Específicas do Cabelo/análise , Masculino , Pessoa de Meia-Idade , Neoplasias Cutâneas/metabolismo , Fatores de Transcrição/análise , Adulto Jovem , beta Catenina/análise
18.
Am J Dermatopathol ; 35(6): 679-84, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23518638

RESUMO

We have recently described a new nail tumor known as onychocytic matricoma. Herein, we describe its malignant counterpart. Clinically, the tumor simulates onychomatricoma (OM). Histologically, this in situ malignant epithelial tumor exhibits a distinct picture of onychocytic differentiation with signs of both nail matrical differentiation and nail plate differentiation. We have proposed the name onychocytic carcinoma for this singular adnexal neoplasm. Given the peculiar thickening of the nail plate observed in OM, onychocytic matricoma, and onychocytic carcinoma, the clinical individualization of a new type of nail band pattern could be proposed. It presents as an acquired localized (monodactylous) longitudinal pachyonychia. Such longitudinal pachyonychia allow the recognition of the matrical nail tumor, which has a limited etiological spectrum. Xantholeucopachyonychia suggests mainly OM and rarely onychocytic carcinoma. Pachymelanonychia suggests onychocytic matricoma and rarely pigmented OM or onychocytic carcinoma.


Assuntos
Carcinoma/classificação , Doenças da Unha/classificação , Unhas/patologia , Neoplasias Cutâneas/classificação , Terminologia como Assunto , Biomarcadores Tumorais/análise , Biópsia , Carcinoma/química , Carcinoma/patologia , Diferenciação Celular , Dedos , Humanos , Imuno-Histoquímica , Queratinas Específicas do Cabelo/análise , Masculino , Pessoa de Meia-Idade , Doenças da Unha/metabolismo , Doenças da Unha/patologia , Unhas/química , Valor Preditivo dos Testes , Neoplasias Cutâneas/química , Neoplasias Cutâneas/patologia
19.
Spectrochim Acta A Mol Biomol Spectrosc ; 109: 226-31, 2013 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-23537511

RESUMO

The effects of scalp dermatitis (seborrheic dermatitis (SD), psoriasis, and atopic dermatitis (AD)) on chemical properties of hair keratin were investigated by Fourier transform infrared (FT-IR) spectroscopy. Hairs were collected from lesional regions affected by SD, psoriasis, and AD and non-lesional regions separately. The hairs with SD were taken from patients with ages of 16-80 years. The ages of patients with psoriasis ranged from 8 to 67 years, and all patients exhibited moderate disease. Hairs with AD were taken from the patients with ages of 24-45 years and the average SCORing atopic dermatitis (SCORAD) was 48.75. Hairs from 20 normal adults were collected as a control. The FT-IR absorbance bands were analyzed by the Gaussian model to obtain the center frequency, half width, height, and area of each band. The height and area of all bands in the spectra were normalized to the amide I centered at 1652 cm(-1) to quantitatively analyze the chemical composition of keratin. The spectra of hair with scalp dermatitis were different with that of control, the amide A components centered at 3278 cm(-1) were smaller than those of the control. The psoriasis hair showed a large difference in the IR absorbance band between lesional and non-lesional hairs indicating good agreement with the morphological changes. The hairs with diseases did not show differences in the content of cystine, which was centered at 1054 cm(-1), from the control. The chemical properties of keratin were not significantly different between the hairs affected by SD, psoriasis, and AD. However, the changes induced by scalp dermatitis were different with weathering. Therefore, FT-IR analysis could be used to screen differences between the physiological and pathological conditions of scalp hair.


Assuntos
Dermatite Atópica/patologia , Dermatite Seborreica/patologia , Cabelo/patologia , Queratinas Específicas do Cabelo/análise , Psoríase/patologia , Couro Cabeludo/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Cabelo/química , Humanos , Masculino , Pessoa de Meia-Idade , Espectroscopia de Infravermelho com Transformada de Fourier , Adulto Jovem
20.
Am J Phys Anthropol ; 150(1): 87-98, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23124593

RESUMO

Historical evidence documents mass migration from Ireland to London during the period of the Great Irish Famine of 1845-52. The rural Irish were reliant on a restricted diet based on potatoes but maize, a C(4) plant, was imported from the United States of America in 1846-47 to mitigate against Famine. In London, Irish migrants joined a population with a more varied diet. To investigate and characterize their diet, carbon and nitrogen isotope ratios were obtained from bone collagen of 119 and hair keratin of six individuals from Lukin Street cemetery, Tower Hamlets (1843-54), and bone collagen of 20 individuals from the cemetery at Kilkenny Union Workhouse in Ireland (1847-51). A comparison of the results with other contemporaneous English populations suggests that Londoners may have elevated δ(15) N compared with their contemporaries in other cities. In comparison, the Irish group have lower δ(15) N. Hair analysis combined with bone collagen allows the reconstruction of perimortem dietary changes. Three children aged 5-15 years from Kilkenny have bone collagen δ(13) C values that indicate consumption of maize (C(4)). As maize was only imported into Ireland in quantity from late 1846 and 1847, these results demonstrate relatively rapid bone collagen turnover in children and highlight the importance of age-related bone turnover rates, and the impact the age of the individual can have on studies of short-term dietary change or recent migration. Stable light isotope data in this study are consistent with the epigraphic and documentary evidence for the presence of migrants within the London cemetery.


Assuntos
Osso e Ossos/química , Migração Humana/história , Inanição/história , Adolescente , Adulto , Osso e Ossos/metabolismo , Isótopos de Carbono/análise , Cemitérios , Criança , Pré-Escolar , Colágeno/análise , Colágeno/química , Interpretação Estatística de Dados , Feminino , História do Século XIX , Humanos , Irlanda/etnologia , Queratinas Específicas do Cabelo/análise , Queratinas Específicas do Cabelo/química , Londres , Masculino , Pessoa de Meia-Idade , Isótopos de Nitrogênio/análise , Sobreviventes
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