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1.
J Biosci ; 44(2)2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31180046

RESUMO

Keratins, the epithelial-predominant members of the intermediate filament superfamily, are expressed in a pairwise, tissuespecific and differentiation-dependent manner. There are 28 type I and 26 type II keratins, which share a common structure comprising a central coiled coil α-helical rod domain flanked by two nonhelical head and tail domains. These domains harbor sites for major posttranslational modifications like phosphorylation and glycosylation, which govern keratin function and dynamics. Apart from providing structural support, keratins regulate various signaling machinery involved in cell growth, motility, apoptosis etc. However, tissue-specific functions of keratins in relation to cell proliferation and differentiation are still emerging. Altered keratin expression pattern during and after malignant transformation is reported to modulate different signaling pathways involved in tumor progression in a context-dependent fashion. The current review focuses on the literature related to the role of keratins in the regulation of cell proliferation, differentiation and transformation in different types of epithelia.


Assuntos
Carcinoma de Células Escamosas/genética , Transformação Celular Neoplásica/genética , Células Epiteliais/metabolismo , Regulação Neoplásica da Expressão Gênica , Queratinas/genética , Neoplasias/genética , Processamento de Proteína Pós-Traducional , Acetilação , Animais , Apoptose/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Diferenciação Celular , Proliferação de Células , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Células Epiteliais/patologia , Glicosilação , Humanos , Queratinas/química , Queratinas/classificação , Queratinas/metabolismo , Neoplasias/metabolismo , Neoplasias/patologia , Fosforilação , Estrutura Secundária de Proteína , Transdução de Sinais
2.
Int. microbiol ; 22(2): 227-237, jun. 2019. graf, tab
Artigo em Inglês | IBECS | ID: ibc-184829

RESUMO

Chicken feather waste is generally insufficiently utilized despite its high content of protein, constituting an environmental issue. Biodegradation of the waste with enabling microbes provides an advantageous option among the available solutions. In this study, an efficient whole feather-degrading strain was strategically isolated from a soil sample taken from a local tea plantation that has little or nothing to do with feathers. The strain was identified as Bacillus thuringiensis (designated as FDB-10) according to the cloned complete 16S rRNA sequence. The FDB-10 could efficiently degrade briefly heat-treated whole feather (102 °C, 5 min; up to 90% of a maximum concentration of 30 g/L) in a salt medium supplemented with 0.1 g/L yeast extract within 24 h (37°C, 150 rpm). Addition of carbon sources (glycerol, glucose, starch, Tween 20, Tween 80, 1.25 g/L as glycerol) to the fermentation medium could improve the degradation. However, significant inhibition could be observed when the added carbon source reached the amount usually adopted in the investigation of carbon source preference (1%). Nitrogen source (NH4Cl, (NH4)2SO4, peptone) adversely influenced the performance of the strain. When the molar concentrations of NH4+ were equal for the two salt, the inhibitory effect on degradation of whole feathers was similar. Entirely different from other reported feather-degrading strains showing a preference to melanin-free feather substrates, the strain isolated in this study could degrade melanin-containing feather equally efficiently, and higher protease activity could be detected in the digest mix. As a plus, the strain could degrade feathers in rice wash produced in daily cooking, indicating its potential use in the simultaneous treatment of rice cooker wastewater produced by a rice processing plant. All these results imply that the FDB-10 is a strain with great potential in the biodegradation of feather waste


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Assuntos
Animais , Bacillus thuringiensis/isolamento & purificação , Plumas/metabolismo , Microbiologia do Solo , Bacillus thuringiensis/genética , Biotransformação , Bacillus thuringiensis/metabolismo , Bacillus thuringiensis/classificação , Galinhas , Análise por Conglomerados , Meios de Cultura , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico , Resíduos Industriais , Queratinas/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Chá/crescimento & desenvolvimento , Temperatura Ambiente
3.
J Nippon Med Sch ; 86(2): 126-130, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31130564

RESUMO

Pseudomyogenic hemangioendothelioma (PMHE) is a new entity. It is an intermediate soft tissue tumor clinically and/or histopathologically mimicking some other high-grade malignant tumors and some inflammatory diseases. We report a case of PMHE on the left plantar surface of a 28-year-old woman. Histopathological examination of the resected specimen revealed spindle and epithelioid cells with plump and atypical nuclei proliferated in the dermis and subcutaneous fat tissue with marked fibroplasia. Both spindle and epithelioid cells had abundant eosinophilic cytoplasm. Neoplastic cells were diffusely positive for AE1/AE3, CK7, vimentin, CD31, FLI-1, ERG, and INI-1. From those findings, we made the diagnosis of PMHE. We describe the main points of differentiation between PMHE and diseases that have similar clinical and/or histopathological findings, including cellular dermatofibroma, spindle cell squamous cell carcinoma, epithelioid sarcoma, epithelioid hemangioendothelioma, epithelioid angiosarcoma, nodular or proliferative fasciitis, and granulomatous fibrosing granulation tissue due to a ruptured epidermal cyst.


Assuntos
Hemangioendotelioma Epitelioide/patologia , Neoplasias de Tecidos Moles/patologia , Adulto , Antiporters/metabolismo , Diagnóstico Diferencial , Feminino , Hemangioendotelioma Epitelioide/diagnóstico , Hemangioendotelioma Epitelioide/genética , Humanos , Queratinas/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Proteína Proto-Oncogênica c-fli-1/metabolismo , Proteína SMARCB1/metabolismo , Neoplasias de Tecidos Moles/diagnóstico , Neoplasias de Tecidos Moles/genética , Regulador Transcricional ERG/metabolismo , Vimentina/metabolismo
4.
BMC Genomics ; 20(1): 411, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31117938

RESUMO

BACKGROUND: Trichophyton rubrum is the main etiological agent of skin and nail infections worldwide. Because of its keratinolytic activity and anthropophilic nature, infection models based on the addition of protein substrates have been employed to assess transcriptional profiles and to elucidate aspects related to host-pathogen interactions. Chalcones are widespread compounds with pronounced activity against dermatophytes. The toxicity of trans-chalcone towards T. rubrum is not fully understood but seems to rely on diverse cellular targets. Within this context, a better understanding of the mode of action of trans-chalcone may help identify new strategies of antifungal therapy and reveal new chemotherapeutic targets. This work aimed to assess the transcriptional profile of T. rubrum grown on different protein sources (keratin or elastin) to mimic natural infection sites and exposed to trans-chalcone in order to elucidate the mechanisms underlying the antifungal activity of trans-chalcone. RESULTS: Overall, the use of different protein sources caused only slight differences in the transcriptional profile of T. rubrum. The main differences were the modulation of proteases and lipases in gene categories when T. rubrum was grown on keratin and elastin, respectively. In addition, some genes encoding heat shock proteins were up-regulated during the growth of T. rubrum on keratin. The transcriptional profile of T. rubrum exposed to trans-chalcone included four main categories: fatty acid and lipid metabolism, overall stress response, cell wall integrity pathway, and alternative energy metabolism. Consistently, T. rubrum Mapk was strongly activated during the first hours of trans-chalcone exposure. Noteworthy, trans-chalcone inhibited genes involved in keratin degradation. The results also showed effects of trans-chalcone on fatty acid synthesis and metabolic pathways involved in acetyl-CoA supply. CONCLUSION: Our results suggest that the mode of action of trans-chalcone is related to pronounced changes in fungal metabolism, including an imbalance between fatty acid synthesis and degradation that interferes with cell membrane and cell wall integrity. In addition, this compound exerts activity against important virulence factors. Taken together, trans-chalcone acts on targets related to dermatophyte physiology and the infection process.


Assuntos
Parede Celular/química , Chalcona/farmacologia , Ácidos Graxos/metabolismo , Proteínas Fúngicas/metabolismo , Tinha/metabolismo , Trichophyton/metabolismo , Fatores de Virulência/antagonistas & inibidores , Antifúngicos/farmacologia , Parede Celular/genética , Elastina/metabolismo , Proteínas Fúngicas/genética , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Humanos , Queratinas/metabolismo , Transdução de Sinais , Tinha/tratamento farmacológico , Tinha/microbiologia , Trichophyton/efeitos dos fármacos , Trichophyton/genética
5.
Genes Cells ; 24(5): 390-402, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30929300

RESUMO

Solo (ARHGEF40) is a RhoA-targeting guanine nucleotide exchange factor that regulates tensional force-induced cytoskeletal reorganization. Solo binds to keratin 8/keratin 18 (K8/K18) filaments through multiple sites, but the roles of these interactions in the localization and mechanotransduction-regulating function of Solo remain unclear. Here, we constructed two Solo mutants (L14R/L17R and L49R/L52R) with leucine-to-arginine replacements in the N-terminal conserved region (which we termed the Solo domain) and analyzed their K18-binding activities. These mutations markedly decreased the K18-binding ability of the N-terminal fragment (residues 1-329) of Solo but had no apparent effect on the K18-binding ability of full-length (FL) Solo. When expressed in cultured cells, wild-type Solo-FL showed a unique punctate localization near the ventral surface of cells and caused the reinforcement of actin filaments. In contrast, despite retaining the K18-binding ability, the L14R/L17R and L49R/L52R mutants of Solo-FL were diffusely distributed in the cytoplasm and barely induced actin cytoskeletal reinforcement. Furthermore, wild-type Solo-FL promoted traction force generation against extracellular matrices and tensional force-induced stress fiber reinforcement, but its L14R/L17R and L49R/L52R mutants did not. These results suggest that the K18-binding ability of the N-terminal Solo domain is critical for the ventral localization of Solo and its function in regulating mechanotransduction.


Assuntos
Fatores de Troca do Nucleotídeo Guanina/metabolismo , Queratinas/metabolismo , Mecanotransdução Celular , Animais , Sítios de Ligação , Cães , Fatores de Troca do Nucleotídeo Guanina/química , Fatores de Troca do Nucleotídeo Guanina/genética , Células HeLa , Humanos , Células Madin Darby de Rim Canino , Mutação , Ligação Proteica
6.
J Sci Food Agric ; 99(11): 4942-4951, 2019 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-30953342

RESUMO

BACKGROUND: To improve the structure-stability and packing characters of collagen fiber, we manufactured crosslinked collagen fiber (CColF)-based edible films using transglutaminase (TGase). Then we made a comparison on structure-stability and packing characteristics among the CColF-based films loaded with casein (CN), keratin (KRT) and soy protein isolate (SPI), respectively. RESULTS: Observed from scanning electron microscopy (SEM), the CColF loaded with CN, KRT and SPI showed some unique morphology of the additional proteins. The CColF-protein films performed better packing characteristics including barrier properties, mechanical properties and thermal-stability properties, compared with CColF films. Importantly, with 500 g kg-1 CN (of CColF) addition, CColF-based films possessed a greater thermal stability than the other films judged from differential scanning calorimetry (DSC). Meanwhile, the CColF loaded with 100 g kg-1 CN provided a higher value of tensile strength (TS) and the CColF loaded with 100 g kg-1 KRT showed a higher value in elongation-at-break (EAB) than the other films. CONCLUSION: In conclusion, the collagen fiber-based edible films with better structure-stability and packing characteristics for food packaging was obtained which could be an advantage to promote the development of the application of collagen in packing products. © 2019 Society of Chemical Industry.


Assuntos
Caseínas/química , Colágeno/química , Reagentes para Ligações Cruzadas , Embalagem de Alimentos/instrumentação , Queratinas/química , Varredura Diferencial de Calorimetria , Caseínas/metabolismo , Reagentes para Ligações Cruzadas/metabolismo , Estabilidade de Medicamentos , Temperatura Alta , Queratinas/metabolismo , Fenômenos Mecânicos , Microscopia Eletrônica de Varredura , Permeabilidade , Proteínas de Soja/química , Proteínas de Soja/metabolismo , Vapor , Resistência à Tração , Transglutaminases/metabolismo , Difração de Raios X
7.
J Vet Med Sci ; 81(6): 821-823, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-30996208

RESUMO

A 12-year-old, male miniature dachshund has an ulcer on the footpad of the right hind limb. Despite treatment for longer than 6 months, the ulcer did not heal. Biopsy of the lesion was done to make a definitive diagnosis. Histologically, there were lumens containing weakly eosinophilic fluid surrounded by tumor cells with a similar circular pale nucleus and distinct nucleoli that showed some variation in size. Immunohistochemically, the tumor cells were positive for cytokeratin (AE1/AE3) and vimentin, were negative for S100 and p63. A poorly differentiated eccrine adenocarcinoma was diagnosed. Treatment was started with toceranib, an anti-angiogenic agent, and enlargement of the lesion was not observed during the administration period.


Assuntos
Adenocarcinoma/veterinária , Doenças do Cão/diagnóstico , Neoplasias das Glândulas Sudoríparas/veterinária , Adenocarcinoma/diagnóstico , Adenocarcinoma/tratamento farmacológico , Inibidores da Angiogênese/uso terapêutico , Animais , Biópsia/veterinária , Doenças do Cão/tratamento farmacológico , Cães , Membro Posterior/patologia , Imuno-Histoquímica , Indóis/uso terapêutico , Queratinas/metabolismo , Masculino , Pirróis/uso terapêutico , Neoplasias das Glândulas Sudoríparas/diagnóstico , Neoplasias das Glândulas Sudoríparas/tratamento farmacológico , Vimentina/metabolismo
8.
J Basic Microbiol ; 59(6): 555-568, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30900760

RESUMO

Keratinases hydrolyze structural protein called keratin into constituent peptides. The present study reports excellent washing efficiency and dehairing properties of thermostable and halotolerant keratinase from Bacillus aerius NSMk2. Alkaline keratinase with molecular mass of 9 kDa displayed remarkable thermostability. K+ , Na+ , Ca2+ , Mn2+ , ß-mercaptoethanol, sodium sulfite, dithiothreitol, ethanol, isopropanol, Tween-20, and Tween-80 stimulated keratinase activity, while Hg2+ and Ba2+ were found to be inhibitory. The enzyme efficiently hydrolyzed a variety of complex protein substrates and exhibited high catalytic efficiency toward keratin-rich substrates and least toward collagen. Keratinase showed exceptional stability to salinity and was found to be compatible with most of the commercial detergents. Efficient removal of chocolate, blood, and egg albumin stains from clothes and tolerance to elevated temperature and salinity potentiated the suitability of keratinase from B. aerius NSMk2 as laundary additive. Keratinase could efficiently dehair goat skin after 15 hr of incubation without damaging the grain structure and collagen layers that assures its use as a promising contender for leather industry.


Assuntos
Bacillus/enzimologia , Proteínas de Bactérias/metabolismo , Detergentes/metabolismo , Microbiologia Industrial , Queratinas/metabolismo , Peptídeo Hidrolases/metabolismo , Bacillus/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Detergentes/química , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Peso Molecular , Peptídeo Hidrolases/química , Peptídeo Hidrolases/isolamento & purificação , Cloreto de Sódio , Especificidade por Substrato , Temperatura Ambiente
9.
Integr Comp Biol ; 59(1): 193-202, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30895301

RESUMO

Corneous proteins are an important component of the tetrapod integument. Duplication and diversification of keratins and associated proteins are linked with the origin of most novel integumentary structures like mammalian hair, avian feathers, and scutes covering turtle shells. Accordingly, the loss of integumentary structures often coincides with the loss of genes encoding keratin and associated proteins. For example, many hair keratins in dolphins and whales have become pseudogenes. The adhesive setae of geckos and anoles are composed of both intermediate filament keratins (IF-keratins, formerly known as alpha-keratins) and corneous beta-proteins (CBPs, formerly known as beta-keratins) and recent whole genome assemblies of two gecko species and an anole uncovered duplications in seta-specific CBPs in each of these lineages. While anoles evolved adhesive toepads just once, there are two competing hypotheses about the origin(s) of digital adhesion in geckos involving either a single origin or multiple origins. Using data from three published gecko genomes, I examine CBP gene evolution in geckos and find support for a hypothesis where CBP gene duplications are associated with the repeated evolution of digital adhesion. Although these results are preliminary, I discuss how additional gecko genome assemblies, combined with phylogenies of keratin and associated protein genes and gene duplication models, can provide rigorous tests of several hypotheses related to gecko CBP evolution. This includes a taxon sampling strategy for sequencing and assembly of gecko genomes that could help resolve competing hypotheses surrounding the origin(s) of digital adhesion.


Assuntos
Evolução Molecular , Duplicação Gênica , Queratinas/genética , Lagartos/fisiologia , Proteínas de Répteis/genética , beta-Queratinas/genética , Animais , Queratinas/metabolismo , Lagartos/genética , Filogenia , Proteínas de Répteis/metabolismo , beta-Queratinas/metabolismo
10.
Biosens Bioelectron ; 131: 104-112, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30826644

RESUMO

Most cancer diagnoses rely on biomarkers detection. This could be improved if directly conducted in suspicious cancer spots, preventing the need for biopsy. Lung cancer remains a perfect study-case for such a development, as it is generally detected at advanced stage and is in the need for early diagnosis techniques. To this aim, we have designed a minimally invasive catheter-embedded biosensor. It combines a specific grating structure photo-imprinted in a telecommunication-grade optical fiber and an overlay made of a thin metal coating on which receptors are grafted, yielding plasmonic coupling. Our optrode targets a type of cytokeratins, overexpressed at the surface of cancer cells. It was assayed ex vivo in resected lung tissues collected from a dozen of patients. Biosensing responses were confirmed by immunohistochemistry, conducted on the same samples. In addition to accurate biosensing, our gratings inherently enable force-sensing features, which also allow a fine positioning of the probe in the tissue. Finally, the in vivo navigation of the bronchoscope-embedded sensor was validated into pig lungs. These achievements are a critical milestone towards the development of this micro/nano biosensor as a cost-effective and weakly invasive diagnostic tool for applications in areas of critical access such as brain, liver or prostate.


Assuntos
Técnicas Biossensoriais , Queratinas/isolamento & purificação , Neoplasias Pulmonares/diagnóstico , Pulmão/metabolismo , Animais , Linhagem Celular Tumoral , Tecnologia de Fibra Óptica , Regulação Neoplásica da Expressão Gênica , Humanos , Queratinas/metabolismo , Pulmão/patologia , Neoplasias Pulmonares/patologia , Fibras Ópticas , Ressonância de Plasmônio de Superfície , Suínos
11.
J Cutan Pathol ; 46(6): 421-424, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30834570

RESUMO

Cutaneous syncytial myoepithelioma (CSM) is a recently recognized, histopathological variant of myoepithelial (ME) tumors of the skin. It is characterized by a syncytial arrangement of spindled, ovoid, and/or epithelioid cells forming a well-circumscribed, unencapsulated dermal nodule. There is a paucity of intervening stroma, and absent duct or gland formation. Strong immunohistochemical staining for S100 and epithelial membrane antigen (EMA) has been described, while cytokeratin expression has been uncommon. The majority of CSMs harbor a rearrangement involving the EWSR1 gene. Although various fusion partner genes have been discovered in ME tumors at other anatomic sites, none has yet been described in CSM. We present a case of CSM represented clinically by a papule on the mid-upper back of a healthy 44-year-old female. It exhibited morphological and immunohistochemical features of a CSM with strong, diffuse S100 and alpha-actin expression, and focal positivity for EMA and cytokeratin AE1/AE3. Fluorescence in-situ hybridization showed an EWSR1 gene rearrangement. Massively parallel next-generation RNA sequencing revealed PBX3 as the fusion partner. The EWSR1-PBX3 gene fusion has been previously identified in three cases of ME tumors of bone and soft tissue, and in a case of retroperitoneal leiomyoma. This is the first report of an EWSR1-PBX3 fusion in CSM.


Assuntos
Biomarcadores Tumorais , Proteínas de Homeodomínio , Mioepitelioma , Proteínas de Fusão Oncogênica , Proteínas Proto-Oncogênicas , Proteína EWS de Ligação a RNA , Neoplasias Cutâneas , Adulto , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Feminino , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Queratinas/genética , Queratinas/metabolismo , Mioepitelioma/genética , Mioepitelioma/metabolismo , Mioepitelioma/patologia , Fusão Oncogênica , Proteínas de Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/metabolismo , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proteína EWS de Ligação a RNA/genética , Proteína EWS de Ligação a RNA/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia
12.
Hum Pathol ; 87: 18-27, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30794893

RESUMO

Tumor buds in colorectal cancer are hypothesized to undergo a (partial) epithelial-mesenchymal transition (EMT). If so, cytokeratin (CK) and vimentin (VIM) co-expression is expected. CK+/VIM+ can also be found in some stromal cells; however, their origin remains unclear. Here, we determine the frequency of CK+/VIM+ tumor cells and characterize the CK+/VIM+ stroma in colorectal cancer. Three cell populations (CK+, VIM+, CK+/VIM+) were sorted using DepArray and fluorescence-activated cell sorting (FACS). Tumor areas were selected to include tumor center, stroma and tumor budding. Fluorescence microscopy was used to visualize co-expressing cells on whole slides. A next-generation tissue microarray (ngTMA) of matched Pan-CK-positive and -negative stroma was constructed and stained for E-cadherin, VIM, Snail1, Twist1, Zeb1 and Zeb2, COL11A1, SPARC, CD90, α-SMA, FAP and WT1. CK+/VIM+ co-expressing tumor cells were detected using all three methods. With DepArray, only tumor budding areas contained CK+/VIM+ cells. The proportion of CK+/VIM+ tumor cells was low (1.5%-22%). CK+ stroma was associated with aggressive tumor features like distant metastasis (P = .0003), lymphatic invasion (P = .0009) and tumor budding (P = .0084). CK+/VIM+ stroma was characterized by positive WT1 (P < .001), ZEB2 (P < .001), TWIST1 (P = .009), and FAP (P = .003). Our data suggest that CK+/VIM+ tumor cells exist, albeit in low numbers and could represent a subgroup of tumor buds in partial EMT. CK+/VIM+ stroma may be of mesothelial origin and shows features of mesenchymal cells and cancer-associated fibroblasts. These results, together with the association with metastasis point to cells in mesothelial-mesenchymal transition (MMT). This atypical stroma may be a potential target for therapy.


Assuntos
Neoplasias Colorretais/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Queratinas/metabolismo , Vimentina/metabolismo , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Humanos , Linfonodos/metabolismo , Linfonodos/patologia , Metástase Linfática/patologia , Masculino , Pessoa de Meia-Idade , Células Estromais/metabolismo , Células Estromais/patologia
13.
Acta Biochim Pol ; 66(1): 111-114, 2019 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-30793712

RESUMO

INTRODUCTION: Small cell lung carcinoma (SCLC) is an aggressive pulmonary neoplasm of neuroendocrine origin. Keratins form a large group of intermediate filaments, which are major structural proteins in epithelial cells and carcinomas. SCLC shows a wide spectrum of keratin expression, from very strong to completely negative. A prognostic role of keratin expression in SCLC is unknown. MATERIAL AND METHODS: Tumor tissue microarray samples from a unique series of 82 SCLC patients who underwent pulmonary resection were stained with keratin specific antibodies AE1/AE3 and CAM5.2. The percentage o1f positively stained cells and their staining pattern (diffusely membranous, partially membranous and dot-like) were evaluated. The median expression value was used for the distinction between keratin-negative and -positive patients. Overall survival in respective groups was compared using the log-rank test. Multivariate Cox proportional hazards regression analysis was performed adjusting for age, gender, tumor site, tumor stage, and tumor histology. RESULTS: edian expression of AE1/AE3 and CAM5.2 was 80% and 90%, respectively. Five cases were completely negative for AE1/AE3 and three for Cam5.2. Median overall survival for patients with stronger and weaker AE1/AE3 staining was 24.7 and 13.8 months, respectively (p=0.019). There was no difference in survival in relation to the CAM5.2 expression (p=0.44). In multivariate analysis adjusted for CAM5.2, T and N stage, gender and age at diagnosis, stronger AE1/AE3 expression was an independent predictor of increased survival (HR 0.50; 95% CI, 0.27-0.94; p=0.031). CONCLUSION: High expression of AE1/AE3 is a favorable prognostic factor in surgically treated SCLC. The applicability of this finding to a typical patient population treated with non-surgical methods warrants further studies.


Assuntos
Queratinas/metabolismo , Carcinoma de Pequenas Células do Pulmão/metabolismo , Antiporters/metabolismo , Biomarcadores/metabolismo , Feminino , Humanos , Pulmão/metabolismo , Pulmão/cirurgia , Masculino , Análise Multivariada , Prognóstico , Modelos de Riscos Proporcionais , Carcinoma de Pequenas Células do Pulmão/patologia , Carcinoma de Pequenas Células do Pulmão/cirurgia , Análise Serial de Tecidos
14.
Int Microbiol ; 22(2): 227-237, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30810985

RESUMO

Chicken feather waste is generally insufficiently utilized despite its high content of protein, constituting an environmental issue. Biodegradation of the waste with enabling microbes provides an advantageous option among the available solutions. In this study, an efficient whole feather-degrading strain was strategically isolated from a soil sample taken from a local tea plantation that has little or nothing to do with feathers. The strain was identified as Bacillus thuringiensis (designated as FDB-10) according to the cloned complete 16S rRNA sequence. The FDB-10 could efficiently degrade briefly heat-treated whole feather (102 °C, 5 min; up to 90% of a maximum concentration of 30 g/L) in a salt medium supplemented with 0.1 g/L yeast extract within 24 h (37 °C, 150 rpm). Addition of carbon sources (glycerol, glucose, starch, Tween 20, Tween 80, 1.25 g/L as glycerol) to the fermentation medium could improve the degradation. However, significant inhibition could be observed when the added carbon source reached the amount usually adopted in the investigation of carbon source preference (1%). Nitrogen source (NH4Cl, (NH4)2SO4, peptone) adversely influenced the performance of the strain. When the molar concentrations of NH4+ were equal for the two salt, the inhibitory effect on degradation of whole feathers was similar. Entirely different from other reported feather-degrading strains showing a preference to melanin-free feather substrates, the strain isolated in this study could degrade melanin-containing feather equally efficiently, and higher protease activity could be detected in the digest mix. As a plus, the strain could degrade feathers in rice wash produced in daily cooking, indicating its potential use in the simultaneous treatment of rice cooker wastewater produced by a rice processing plant. All these results imply that the FDB-10 is a strain with great potential in the biodegradation of feather waste.


Assuntos
Bacillus thuringiensis/isolamento & purificação , Bacillus thuringiensis/metabolismo , Plumas/metabolismo , Microbiologia do Solo , Animais , Bacillus thuringiensis/classificação , Bacillus thuringiensis/genética , Biotransformação , Galinhas , Análise por Conglomerados , Meios de Cultura , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fermentação , Resíduos Industriais , Queratinas/metabolismo , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Chá/crescimento & desenvolvimento , Temperatura Ambiente
16.
J Korean Med Sci ; 34(1): e5, 2019 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-30618513

RESUMO

Background: Cutaneous carcinosarcoma is a rare biphasic tumor comprising malignant epithelial and heterologous mesenchymal elements. Data on the clinical and histopathologic characteristics of this tumor in Asian populations are not available. The purpose of this study was to investigate the clinicopathologic and immunohistochemical features of cutaneous carcinosarcoma in the Korean population. Methods: We retrospectively reviewed the records of 11 patients with cutaneous carcinosarcoma who were diagnosed from 2006 to 2016. Results: The mean patient age at diagnosis was 71.5 years (range, 43-96 years) and there was a men predilection. The most common site of cutaneous carcinosarcoma was the head and neck (8/11, 72.7%). Histopathologically, most tumors showed a characteristic morphology consisting of two types of tumor cells, varied differentiated epithelial cells (such as basal or squamous cells) and spindle cells with transition zones between the two components. These two cell types also demonstrated variable immunohistochemical characteristics. Conclusion: Although the number of cases in this study was limited, our results provide valuable insight into the clinical and histopathologic characteristics of cutaneous carcinosarcoma in the Korean population.


Assuntos
Carcinossarcoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Basocelular/metabolismo , Carcinoma Basocelular/patologia , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Carcinossarcoma/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Queratinas/metabolismo , Masculino , Pessoa de Meia-Idade , República da Coreia , Estudos Retrospectivos , Proteína Supressora de Tumor p53/metabolismo
17.
Mater Sci Eng C Mater Biol Appl ; 96: 616-624, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30606573

RESUMO

Intermediate filaments, together with actin microfilaments and microtubules constituent the cytoskeleton of mammalian cells, involving in various cellular activities. The roles of intermediate filaments in cell skeleton reorganization when responding with extracellular matrix (ECM) nanostructure are poorly understood yet. To unveil the effects of fibrous composition and orientation on cells, we developed electrospun nanofibers of varying topology and components, and the effects on assembly of intermediate filaments as keratin and vimentin were investigated in detail. We found that aligned nanofibers enhanced expression of E-cadherin and promoted assembly of keratin intermediate filaments. Meanwhile, the compositional variation show different preference on up-regulation of the two intermediate filaments. Compared to keratin, the assembly of vimentin intermediate filaments were promoted by incorporating bovine serum albumin (BSA) functionalized graphene oxide (BSA-GO) into polycaprolactone (PCL) nanofibers. Thus, our findings elucidate how the different physical factors of fibrous extracellular matrix affect the reorganization of cytoskeleton by assembly of keratin and vimentin intermediate filaments.


Assuntos
Matriz Extracelular/química , Filamentos Intermediários/metabolismo , Queratinas/metabolismo , Nanofibras/química , Proteínas de Neoplasias/metabolismo , Neoplasias Pancreáticas/metabolismo , Vimentina/metabolismo , Linhagem Celular Tumoral , Humanos , Filamentos Intermediários/patologia , Neoplasias Pancreáticas/patologia
18.
Mol Phylogenet Evol ; 133: 352-361, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30599197

RESUMO

Mammalian genomes contain a number of duplicated genes, and sequence identity between these duplicates can be maintained by purifying selection. However, between-duplicate recombination can also maintain sequence identity between copies, resulting in a pattern known as concerted evolution where within-genome repeats are more similar to each other than to orthologous repeats in related species. Here we investigated the tandemly-repeated keratin-associated protein 1 (KAP1) gene family, KRTAP1, which encodes proteins that are important components of hair and wool in mammals. Comparison of eutherian mammal KRTAP1 gene repeats within and between species shows a strong pattern of concerted evolution. However, in striking contrast to the coding regions of these genes, we find that the flanking regions have a divergent pattern of evolution. This contrast in evolutionary pattern transitions abruptly near the start and stop codons of the KRTAP1 genes. We reveal that this difference in evolutionary patterns is not explained by conventional purifying selection, nor is it likely a consequence of codon adaptation or reverse transcription of KRTAP1-n mRNA. Instead, the evidence suggests that these contrasting patterns result from short-tract gene conversion events that are biased to the KRTAP1 coding region by selection and/or differential sequence divergence. This work demonstrates the power that gene conversion has to finely shape the evolution of repetitive genes, and provides another distinctive pattern of contrasting evolutionary outcomes that results from gene conversion. A greater emphasis on exploring the evolution of multi-gene eukaryotic families will reveal how common different contrasting evolutionary patterns are in gene duplicates.


Assuntos
Evolução Molecular , Queratinas/genética , Mamíferos/genética , Fases de Leitura Aberta/genética , Animais , Sequência de Bases , Códon/genética , DNA Intergênico/genética , Conversão Gênica , Queratinas/metabolismo , Filogenia , Polimorfismo Genético , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Seleção Genética , Ovinos/genética , Sequências de Repetição em Tandem/genética
19.
PLoS One ; 14(1): e0210504, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30699132

RESUMO

ORF virus (ORFV) is the causative agent of contagious ecthyma, a pustular dermatitis of small ruminants and humans. Even though the development of lesions caused by ORFV was extensively studied in animals, only limited knowledge exists about the lesion development in human skin. The aim of the present study was to evaluate a three-dimensional (3D) organotypic culture (OTC) as a human skin model for ORFV infection considering lesion development, replication of the virus, viral gene transcription and modulation of differentiation of human keratinocytes by ORFV. ORFV infection of OTC was performed using the ORFV isolate B029 derived from a human patient. The OTC sections showed a similar structure of stratified epidermal keratinocytes as human foreskin and a similar expression profile of the differentiation markers keratin 1 (K1), K10, and loricrin. Upon ORFV infection, OTCs exhibited histological cytopathic changes including hyperkeratosis and ballooning degeneration of the keratinocytes. ORFV persisted for 10 days and was located in keratinocytes of the outer epidermal layers. ORFV-specific early, intermediate and late genes were transcribed, but limited viral spread and restricted cell infection were noticed. ORFV infection resulted in downregulation of K1, K10, and loricrin at the transcriptional level without affecting proliferation as shown by PCNA or Ki-67 expression. In conclusion, OTC provides a suitable model to study the interaction of virus with human keratinocytes in a similar structural setting as human skin and reveals that ORFV infection downregulates several differentiation markers in the epidermis of the human skin, a hitherto unknown feature of dermal ORFV infection in man.


Assuntos
Diferenciação Celular , Ectima Contagioso/virologia , Prepúcio do Pênis/virologia , Queratinócitos/virologia , Vírus do Orf/fisiologia , Técnicas de Cultura de Órgãos/métodos , Animais , Linhagem Celular , Células Cultivadas , Ectima Contagioso/genética , Ectima Contagioso/metabolismo , Prepúcio do Pênis/crescimento & desenvolvimento , Prepúcio do Pênis/metabolismo , Perfilação da Expressão Gênica , Humanos , Queratinócitos/metabolismo , Queratinócitos/patologia , Queratinas/genética , Queratinas/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Organogênese , Ovinos
20.
Eur J Pharmacol ; 846: 86-99, 2019 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-30641059

RESUMO

Many ent-kaurane diterpenoids exhibit notable antitumor activity in vitro and in vivo, and some have been used as cancer therapeutic agents in China. In this study, we identified a novel molecular target of leukamenin E, an ent-kaurane diterpenoid, using an available whole-cell model in combination with immunofluorescence imaging and mass spectrometry (MS). The cytoskeleton-disrupting drugs cytochalasin B and colchicine caused the depolymerization of microfilaments and the collapse of microtubules and vimentin filaments, respectively, but had little effects on HepG2 and NCI-H1299 cells spreading as well as keratin filament (KF) reassembly, indicating that KFs are involved in cell spreading. Leukamenin E blocked HepG2 and NCI-H1299 cells adhesion/spreading and KF reassembly at subtoxic concentrations, indicating that leukamenin E may target KFs. Moreover, leukamenin E, at 3 µM for 24 h or 10 µM for 3 h, induced massive KF depolymerization in well-spread HepG2 and NCI-H1299 cells treated with/without cytochalasin B and colchicine. MS analysis indicated that leukamenin E could covalently modify amino acid residue(s) in a synthetic peptide based on keratin 1 and keratin 10 sequences, suggesting that covalent modification of the synthetic peptide by leukamenin E caused assembly inhibition or disrupted KF polymerization in HepG2 and NCI-H1299 cells. In addition, acridine orange/ethidium bromide staining and western blotting confirmed that there was no correlation between the KF-disrupting effects and apoptosis or keratin expression. Thus, we propose that leukamenin E is a novel inhibitor of KF assembly, and as such, can serve as a chemical probe of KF functions and a potential molecular target for ent-kaurane diterpenoid-based therapeutics.


Assuntos
Citoesqueleto de Actina/metabolismo , Adesão Celular/efeitos dos fármacos , Diterpenos de Caurano/farmacologia , Queratinas/metabolismo , Fatores de Despolimerização de Actina , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Colchicina/farmacologia , Citocalasina B/farmacologia , Células Hep G2 , Humanos , Espectrometria de Massas/métodos , Microscopia de Fluorescência/métodos , Microtúbulos/efeitos dos fármacos , Vimentina/metabolismo
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