Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 364
Filtrar
1.
Anticancer Res ; 39(12): 6567-6573, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31810922

RESUMO

BACKGROUND/AIM: The KIAA1199 gene has been associated with cancer-cell proliferation, but its functions remain poorly studied. Here, we examined the clinical significance of the KIAA1199 mRNA levels in locally advanced gastric cancer (GC). Materials and Methods/Results: Using samples from 254 patients with stage II/III GC, we found significantly higher KIAA1199 levels in cancerous tissues compared to adjacent normal mucosa (ANM). There was no significant relationship between KIAA1199 expression and clinical features. Although overall survival rates (OSR) of patients, who underwent surgery did not correlate with KIAA1199 expression, patients who underwent adjuvant chemotherapy with S-1 and had high KIAA1199 levels displayed significantly lower OSR. KIAA1199 knock down (KIAA1199-KD) suppressed proliferation, invasiveness, and sensitivity of GC cells to 5-fluorouracil (5-FU). CONCLUSION: KIAA1199 expression appears to be a promising prognostic marker in patients with locally advanced GC, who underwent postoperative adjuvant chemotherapy with S-1. KIAA1199 may represent a novel target for GC pharmacotherapy.


Assuntos
Hialuronoglucosaminidase/genética , Ácido Oxônico/uso terapêutico , Neoplasias Gástricas/terapia , Tegafur/uso terapêutico , Regulação para Cima , Idoso , Linhagem Celular Tumoral , Proliferação de Células , Quimioterapia Adjuvante , Procedimentos Cirúrgicos do Sistema Digestório , Combinação de Medicamentos , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Prognóstico , Neoplasias Gástricas/genética , Neoplasias Gástricas/mortalidade , Análise de Sobrevida
2.
Vet J ; 254: 105393, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31836163

RESUMO

Mammary gland tumors are a heterogeneous group of neoplastic diseases. Genetic studies make it possible to determine genetic profiles and identify new molecular markers. The aim of the study was to evaluate the gene expression profile of canine mammary carcinomas and identify potential prognostic markers. Twelve mammary cancer samples from bitches were collected for the evaluation of global gene expression. Microarray assays were performed using commercial kits. Statistical analysis of the microarray was done using moderate t-statistic and adjusted using the Benjamini and Hochberg procedure. Differential connectivity analysis was also performed. Enrichment analyses were conducted using WebGestalt. P-values were calculated using hypergeometric statistics and adjusted using the Benjamini and Hochberg procedure. The HYAL-1 gene was validated using quantitative PCR (qPCR). There were 878 upregulated genes and 821 downregulated genes in the neoplasms studied. Enrichment analysis (individual analysis) identified the HYAL-1 gene as a potential marker of tumorigenesis and tumor recurrence. Differential connectivity analysis demonstrated 262 differentially connected genes.


Assuntos
Doenças do Cão/genética , Neoplasias Mamárias Animais/genética , Animais , Biomarcadores Tumorais/metabolismo , DNA de Neoplasias , Doenças do Cão/enzimologia , Cães , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Neoplasias Mamárias Animais/enzimologia , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real/veterinária
3.
Nat Cell Biol ; 21(11): 1403-1412, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31685984

RESUMO

The development of effective therapies against brain metastasis is currently hindered by limitations in our understanding of the molecular mechanisms driving it. Here we define the contributions of tumour-secreted exosomes to brain metastatic colonization and demonstrate that pre-conditioning the brain microenvironment with exosomes from brain metastatic cells enhances cancer cell outgrowth. Proteomic analysis identified cell migration-inducing and hyaluronan-binding protein (CEMIP) as elevated in exosomes from brain metastatic but not lung or bone metastatic cells. CEMIP depletion in tumour cells impaired brain metastasis, disrupting invasion and tumour cell association with the brain vasculature, phenotypes rescued by pre-conditioning the brain microenvironment with CEMIP+ exosomes. Moreover, uptake of CEMIP+ exosomes by brain endothelial and microglial cells induced endothelial cell branching and inflammation in the perivascular niche by upregulating the pro-inflammatory cytokines encoded by Ptgs2, Tnf and Ccl/Cxcl, known to promote brain vascular remodelling and metastasis. CEMIP was elevated in tumour tissues and exosomes from patients with brain metastasis and predicted brain metastasis progression and patient survival. Collectively, our findings suggest that targeting exosomal CEMIP could constitute a future avenue for the prevention and treatment of brain metastasis.


Assuntos
Neoplasias Encefálicas/genética , Exossomos/metabolismo , Regulação Neoplásica da Expressão Gênica , Hialuronoglucosaminidase/genética , Neovascularização Patológica/genética , Microambiente Tumoral/genética , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Quimiocina CCL1/genética , Quimiocina CCL1/metabolismo , Quimiocina CXCL1/genética , Quimiocina CXCL1/metabolismo , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Exossomos/patologia , Humanos , Hialuronoglucosaminidase/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Metástase Neoplásica , Neovascularização Patológica/metabolismo , Neovascularização Patológica/mortalidade , Neovascularização Patológica/patologia , Transdução de Sinais , Análise de Sobrevida , Carga Tumoral , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
4.
Med Sci Monit ; 25: 6788-6796, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31501407

RESUMO

BACKGROUND KIAA1199 has been reported to be associated with malignant progression and poor clinical outcomes in various human malignancies. However, its clinical role and molecular function remain unknown in papillary thyroid cancer (PTC). MATERIAL AND METHODS The Cancer Genome Atlas (TCGA) was used to investigate the expression profiles of KIAA1199 and miR-486-5p in PTC. Immunohistochemistry was used to validate the protein expression of KIAA1199 in PTC. The Weighted Gene Co-expression Network Analysis (WGCNA) and Gene Set Enrichment Analysis (GSEA) were used to explore the potential pathway underling KIAA1199 in PTC. In vitro and in vivo experiments were performed to investigate the biological role of KIAA1199 in PTC progression. Luciferase reporter assays and Western blot analysis were performed to determine whether KIAA1199 is a downstream target of miR-486-5p. RESULTS We found that KIAA1199 was aberrantly elevated in PTC tissues compared with normal tissues, and upregulation of KIAA1199 was positively correlated with more advanced clinical variables. Additionally, bioinformatic analysis indicated that KIAA1199 was involved in cell migration and invasion. KIAA1199 silencing inhibited the invasive ability of PTC cells by affecting epithelial-mesenchymal transition (EMT) in vitro and in vivo. Furthermore, miR-486-5p was identified as an upstream microRNA that directly targets the 3'-UTR region of KIAA1199. CONCLUSIONS The miR-486-5p/KIAA1199/EMT axis might play a critical role in PTC invasion and metastasis and offers a potential therapeutic strategy for PTC.


Assuntos
Transição Epitelial-Mesenquimal/genética , Hialuronoglucosaminidase/metabolismo , MicroRNAs/metabolismo , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/patologia , Animais , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Hialuronoglucosaminidase/genética , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Invasividade Neoplásica , Regulação para Cima/genética
5.
Oncol Rep ; 42(5): 2065-2074, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31545463

RESUMO

O­linked ß­N­acetylglucosamine (O­GlcNAc) modification is a dynamic post­translational modification process that is involved in many crucial biological processes, including cell cycle regulation, nutrient metabolism and extracellular signaling. This dynamic modification is dependent on the ambient glucose concentration and is catalyzed and removed by O­GlcNAc transferase (OGT) and O­GlcNAcase (OGA), respectively. The present study aimed to determine the role of O­GlcNAcylation during embryo implantation by inhibiting or enhancing its function and expression. The results revealed that the expression of O­GlcNAc­modified proteins in the human secretory endometrium was higher than that of the endometrium during the proliferative phase, as determined via western blotting and immunohistochemistry. Additionally, the level of endometrial O­GlcNAc modification increased gradually from the pre­receptive to the receptive phase, which was then decreased during the non­receptive phase. In endometrial cells, RNA interference was utilized to reduce the expression of two key O­GlcNAc synthesis and decomposition enzymes, OGT and OGA, to indirectly increase or decrease levels of O­GlcNAc modification. The results revealed that increasing the level of O­GlcNAc modification enhanced cellular proliferation, migration, invasion and adhesion, thereby promoting embryo implantation. It is hypothesized that O­GlcNAc modification serves an important role in the regulation of endometrial receptivity and embryo implantation. The results of the present study may have important implications for the understanding of female fertility and may help improve infertility treatments.


Assuntos
Acetilglucosamina/metabolismo , Antígenos de Neoplasias/metabolismo , Endométrio/metabolismo , Histona Acetiltransferases/metabolismo , Hialuronoglucosaminidase/metabolismo , N-Acetilglucosaminiltransferases/metabolismo , Adulto , Antígenos de Neoplasias/genética , Linhagem Celular , Movimento Celular , Proliferação de Células , Implantação do Embrião , Feminino , Fase Folicular/metabolismo , Glicosilação , Histona Acetiltransferases/genética , Humanos , Hialuronoglucosaminidase/genética , Pessoa de Meia-Idade , N-Acetilglucosaminiltransferases/genética
6.
J Microbiol Biotechnol ; 29(8): 1310-1315, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31370115

RESUMO

Hyaluronidases enhance therapeutic drug transport by breaking down the hyaluronan barrier to lymphatic and capillary vessels, facilitating their tissue absorption. Commercially available hyaluronidases are bovine in origin; however, they pose risks such as bovine spongiform encephalopathy. The present study aimed to develop a novel, highly active hyaluronidase and assess its function. Therefore, in order to find the most efficient active hyaluronidase, we produced several shortened hyaluronidases with partial removal of the N- or C-terminal regions. Moreover, we created an enzyme that connected six histidines onto the end of the hyaluronidase C-terminus. This simplified subsequent purification using Ni2+ affinity chromatography, making it feasible to industrialize this highly active recombinant hyaluronidase which exhibited catalytic activity equal to that of the commercial enzyme. Therefore, this simple and effective isolation method could increase the availability of recombinant hyaluronidase for research and clinical purposes.


Assuntos
Histidina/metabolismo , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Oligopeptídeos/metabolismo , Proteínas Recombinantes , Animais , Bovinos , Moléculas de Adesão Celular/metabolismo , Clonagem Molecular , Estabilidade Enzimática , Células HEK293 , Humanos , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/isolamento & purificação , Concentração de Íons de Hidrogênio , Temperatura Ambiente
7.
PLoS One ; 14(7): e0218736, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31260471

RESUMO

LL-37 is the only human cathelicidin-family host defense peptide and has been reported to interact with invading pathogens causing inflammation at various body sites. Recent studies showed high levels of LL-37 in the synovial-lining membrane of patients with rheumatoid arthritis, a common type of inflammatory arthritis. The present study aims to investigate the role of LL-37 on mechanisms associated with pathogenesis of inflammatory arthritis. The effects of LL-37 on the expression of proinflammatory cytokines, hyaluronan (HA) metabolism-related genes, cell death-related pathways, and cell invasion were investigated in SW982, a human synovial sarcoma cell line. Time-course measurements of proinflammatory cytokines and mediators showed that LL-37 significantly induced IL6 and IL17A mRNA levels at early time points (3-6 hr). HA-metabolism-related genes (i.e., HA synthase 2 (HAS2), HAS3, hyaluronidase 1 (HYAL1), HYAL2, and CD44) were co-expressed in parallel. In combination, LL-37 and IL17A significantly enhanced PTGS2, TNF, and HAS3 gene expression concomitantly with the elevation of their respective products, PGE2, TNF, and HA. Cell invasion rates and FN1 gene expression were also significantly enhanced. However, LL-37 alone or combined with IL17A did not affect cell mortality or cell cycle. Treatment of SW982 cells with both LL-37 and IL17A significantly enhanced IKK and p65 phosphorylation. These findings suggest that the chronic production of a high level of LL-37 may synchronize with its downstream proinflammatory cytokines, especially IL17A, contributing to the co-operative enhancement of pathogenesis mechanisms of inflammatory arthritis, such as high production of proinflammatory cytokines and mediators together with the activation of HA-metabolism-associated genes and cell invasion.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Ácido Hialurônico/metabolismo , Interleucina-17/farmacologia , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/imunologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Combinação de Medicamentos , Sinergismo Farmacológico , Fibroblastos/imunologia , Fibroblastos/patologia , Fibronectinas/genética , Fibronectinas/imunologia , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/imunologia , Regulação da Expressão Gênica/imunologia , Humanos , Receptores de Hialuronatos/genética , Receptores de Hialuronatos/imunologia , Hialuronan Sintases/genética , Hialuronan Sintases/imunologia , Ácido Hialurônico/imunologia , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/imunologia , Quinase I-kappa B/genética , Quinase I-kappa B/imunologia , Inflamação , Interleucina-6/genética , Interleucina-6/imunologia , Transdução de Sinais , Membrana Sinovial/imunologia , Membrana Sinovial/patologia , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
8.
Int Immunopharmacol ; 73: 203-211, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31103876

RESUMO

The overproduction of proteolytic enzymes and dysregulation of extracellular matrix (ECM) metabolism have been shown to accelerate the degradation process of articular cartilage. The purpose of this study was to investigate the role of KIAA1199 and its association with the pathophysiology of osteoarthritis (OA). We found that the expression of KIAA1199 was significantly upregulated in OA cartilage compared with normal tissues. Serum levels of KIAA1199 were higher in OA patients than in non-OA patients. Furthermore, knockdown of KIAA1199 inhibited interleukin-1 beta (IL-1ß)-induced ECM metabolic imbalance by regulating the expression of A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 5; matrix metallopeptidase-13; aggrecan; and COL2A1. In addition, silencing of KIAA1199 significantly decreased the expression of inflammatory mediators such as prostaglandin E2, IL-6, and TNF-α. Mechanistic analyses further revealed that IL-1ß-induced activation of the Wnt/ß-catenin pathway was suppressed during KIAA1199 knockdown. Moreover, KIAA1199 expression was also upregulated in an in vivo rat OA model. Together, these results increase our understanding of the emerging role of KIAA1199 in the process of OA degeneration, and may lead to a novel molecular target to prevent cartilage degradation.


Assuntos
Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Hialuronoglucosaminidase/metabolismo , Interleucina-1beta , Osteoartrite/metabolismo , Via de Sinalização Wnt , Animais , Células Cultivadas , Técnicas de Silenciamento de Genes , Humanos , Hialuronoglucosaminidase/sangue , Hialuronoglucosaminidase/genética , Masculino , Ratos Sprague-Dawley , Regulação para Cima
9.
J Agric Food Chem ; 67(18): 5240-5249, 2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31008594

RESUMO

Fluoride is a widespread environmental pollutant that can induce low sperm quality and fertilizing ability; however, the underlying mechanism still remains unclear. Hence, we aimed to investigate the influence of fluoride on the sperm fertilizing ability via some key proteins in the epididymis. For this, 40 adult rats were assigned randomly into four groups. The control group was given distilled water, while the other three groups were given 25, 50, and 100 mg of NaF/L via drinking water for 56 days, respectively. After 1 day, epididymides were processed for sperm-egg binding, RNA extraction, western blot, and immunofluorescence analysis. Fluoride exposure reduced the ability of sperm to break down the egg cumulus cell layer. A further study revealed that fluoride altered the expression levels of genes and proteins related to acrosome reaction in vivo, including SPAM1, ACR, and PRSS21. However, fluoride only affected the expression of the ACR protein only in the epididymis but not in the testis. Fluoride also affected the expression levels of the membrane proteins CD9 and CD81 of epididymosomes in the epididymis. From the results, it can be concluded that fluoride exposure reduced the ability of sperm to break down the egg cumulus cell layer, which could be one of the reasons for decreased fertility ability in males treated with fluoride. These results provide some theoretical guidance and new ideas for treatments of low fertility, infertility, and other reproductive diseases.


Assuntos
Acrosina/metabolismo , Moléculas de Adesão Celular/metabolismo , Epididimo/efeitos dos fármacos , Fluoretos/farmacologia , Hialuronoglucosaminidase/metabolismo , Serina Endopeptidases/metabolismo , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Acrosina/genética , Animais , Moléculas de Adesão Celular/genética , Epididimo/metabolismo , Fertilização/efeitos dos fármacos , Hialuronoglucosaminidase/genética , Masculino , Ratos , Ratos Sprague-Dawley , Serina Endopeptidases/genética , Testículo/efeitos dos fármacos , Testículo/metabolismo
10.
Biochimie ; 162: 33-45, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30946947

RESUMO

Snake venoms are a rich source of enzymes such as metalloproteinases, serine proteinases phospholipases A2 and myotoxins, that have been well characterized structurally and functionally. However, hyaluronidases (E.C.3.2.1.35) have not been studied extensively. In this study, we describe the biochemical and molecular features of a hyaluronidase (Hyal-Ba) isolated from the venom of the Peruvian snake Bothrops atrox. Hyal-Ba was purified by a combination of ion-exchange and gel filtration chromatography. Purified Hyal-Ba is a 69-kDa (SDS-PAGE) monomeric glycoprotein with an N-terminal amino acid sequence sharing high identity with homologous snake venom hyaluronidases. Detected associated carbohydrates were hexoses (16.38%), hexosamines (2.7%) and sialic acid (0.69%). Hyal-Ba selectively hydrolyzed only hyaluronic acid (HA; specific activity = 437.5 U/mg) but it did not hydrolyze chondroitin sulfate or heparin. The optimal pH and temperature for maximum activity were 6.0 and 40 °C, respectively, and its Km was 0.31 µM. Its activity was inhibited by EDTA, iodoacetate, 2-mercaptoethanol, TLCK and dexamethasone. Na+ and K+ (0.2 M) positively affect hyaluronidase activity; while Mg2+, Br2+, Ba2+, Cu2+, Zn2+, and Cd2+ reduced catalytic activity. Hyal-Ba potentiates the hemorrhagic and hemolytic activity of whole venom, but decreased subplantar edema caused by an l-amino acid oxidase (LAAO). The Hyal-Ba cDNA sequence (2020 bp) encodes 449 amino acid residues, including the catalytic site residues (Glu135, Asp133, Tyr206, Tyr253 and Trp328) and three functional motifs for N-linked glycosylation, which are conserved with other snake hyaluronidases. Spatial modeling of Hyal-Ba displayed a TIM-Barrel (α/ß) fold and an EGF-like domain in the C-terminal portion. The phylogenetic analysis of Hyal-Ba with other homologous Hyals showed the monophyly of viperids. Further, Hyal-Ba studies may extend our knowledge of B. atrox toxinology and provides insight to improve the neutralizing strategies of therapeutic antivenoms.


Assuntos
Bothrops/metabolismo , Venenos de Crotalídeos , Hialuronoglucosaminidase , Animais , Sequência de Bases/genética , Venenos de Crotalídeos/enzimologia , Venenos de Crotalídeos/toxicidade , DNA Complementar , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/classificação , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/toxicidade , Cinética , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Peru , Filogenia , Estabilidade Proteica , Estrutura Secundária de Proteína , Especificidade por Substrato
11.
Virchows Arch ; 475(2): 251-254, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30911815

RESUMO

Pleomorphic hyalinizing angiectatic tumor (PHAT) is a rare mesenchymal tumor of intermediate malignancy. PHAT, and the related hemosiderotic fibrolipomatous tumor, show a recurrent t(1;10)(p22;q24). Fluorescence in situ hybridization (FISH) and BAC (bacterial artificial chromosome) clones have previously identified TGFBR3 and MGEA5 as fusion partners. However, targeted RNA-sequencing allowed for the correct identification of FBXW4 and not MGEA5 as the fusion partner of TGFBR3 in a subcutaneous PHAT, a finding further confirmed by RT-PCR. FBXW4 and MGEA5 share a common cytogenetic location at 10q24.32, thereby suggesting that the use of less precise technology may have led to inaccurate gene identification. The study of additional cases is however required.


Assuntos
Antígenos de Neoplasias/genética , Histona Acetiltransferases/genética , Hialuronoglucosaminidase/genética , Proteoglicanas/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Sarcoma/genética , Análise de Sequência de RNA/métodos , Neoplasias de Tecidos Moles/genética , Proteínas F-Box/genética , Humanos , Masculino , Pessoa de Meia-Idade , Fusão Oncogênica , Sarcoma/patologia , Neoplasias de Tecidos Moles/patologia
12.
Behav Brain Res ; 364: 183-192, 2019 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-30738099

RESUMO

Recent genome-wide association study (GWAS) identified 12 independent loci for Attention-deficit hyperactivity disorder (ADHD). However, the causal genes expression and pathways of ADHD is still vague. We integrated GWAS, eQTL and genes expression data to find the causal genes, genes expression, and genes prioritization in the different brain tissues and whole blood cells. Overall 47 genes were prioritized, the most promising genes were LSG1, HYAL3, PIDD, PNPLA2, BLOC1S2, PLK1S1, CALN1, KAT2B, CTNNB1 and WDR11. Whereas, the CALN1, KAT2B, and WDR11 were previously associated with schizophrenia (SZ), bipolar (BP) and drug abuse. Gene ontology analysis shows that the glutamate receptor signaling pathway (P = 8.009E-07, with false discovery rate (FDR) < 5%), GRIK5 sub network (P = 2.887E-06, FDR < 5%), abnormal gait (P = 3.657E-06, FDR < 5%), REACTOME_SIGNALING_BY_ERBB2 (P = 5.161E-06, FDR < 5%), and abnormal nervous system physiology (P = 5.239E-06, FDR < 5%) were associated with ADHD. These causal genes were highly expressed in Fetal Astrocytes, Neurons, and Microglia/Macrophage. This study illustrates the comprehensive GWAS integrative approach of ADHD. However, further genetic and functional studies are required to validate the role of these genes in the etiology of ADHD, which should provide novel insights into the understanding of this disease.


Assuntos
Transtorno do Deficit de Atenção com Hiperatividade/genética , Mapeamento Encefálico/métodos , Transtorno do Deficit de Atenção com Hiperatividade/fisiopatologia , Encéfalo/metabolismo , Moléculas de Adesão Celular/genética , Bases de Dados Genéticas , Proteínas Adaptadoras de Sinalização de Receptores de Domínio de Morte/genética , GTP Fosfo-Hidrolases/genética , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla , Humanos , Hialuronoglucosaminidase/genética , Lipase/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Polimorfismo de Nucleotídeo Único/genética , Mapas de Interação de Proteínas/genética , Locos de Características Quantitativas/genética , Transcriptoma/genética
13.
Adv Mater ; 31(34): e1803549, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30773699

RESUMO

There is a rapidly increasing interest in developing stimuli-responsive nanomaterials for treating a variety of diseases. By enabling the activation of function locally at the sites of interest, it is possible to increase therapeutic efficacy significantly while simultaneously reducing adverse side effects. While there are many sophisticated nanomaterials available, they are often highly complex and not easily transferrable to industrial scales and clinical settings. However, nanomaterials based on hyaluronic acid offer a compelling strategy for reducing their complexity while retaining several desirable benefits such as active targeting and stimuli-responsive degradation. Herein, the basic properties of hyaluronic acid, its binding partners, and natural routes for degradation by hyaluronidases-hyaluronic-acid-degrading enzymes-and oxidative stresses are discussed. Recent advances in designing hyaluronic acid-based, actively targeted, hyaluronidase- or reactive-oxygen-species-responsive nanomaterials for both diagnostic imaging and therapeutic delivery, which go beyond merely the classical targeting of CD44, are summarized.


Assuntos
Portadores de Fármacos/química , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/química , Nanoestruturas/química , Animais , Diagnóstico por Imagem , Técnicas de Transferência de Genes , Humanos , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Imagem Molecular , Terapia de Alvo Molecular , Estresse Oxidativo , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo
14.
Arch Pathol Lab Med ; 143(2): 212-221, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-29979612

RESUMO

CONTEXT.­: Pleomorphic hyalinizing angiectatic tumor (PHAT) of soft parts, hemosiderotic fibrolipomatous tumor (HFLT), and myxoinflammatory fibroblastic sarcoma (MIFS) are 3 distinct entities of low-grade spindle cell mesenchymal neoplasm. These tumors have similar clinical presentations and partially overlapping but distinctive pathologic features. A recurrent translocation, t(1;10)(p22;q24), has been detected in a subset of PHAT, HFLT, MIFS, and HFLT/MIFS hybrid cases. Translocation t(1;10)(p22;q24) involves transforming growth factor ß-receptor 3 ( TGFBR3) and meningioma-expressed antigen 5 ( MGEA5) genes on chromosomes 1p22 and 10q24, respectively. However, the percentage of translocation in PHAT, HFLT, and MIFS varies significantly among different studies. The relationship among these tumors has been a controversial topic among experts. OBJECTIVE.­: To discuss the diagnostic and functional significance of translocation t(1;10)(p22;q24) TGFBR3/MGEA5 rearrangement in HFLT, PHAT, and MIFS. DATA SOURCES.­: PubMed was used for this study. CONCLUSIONS.­: Diagnosis of HFLT, PHAT, and MIFS is challenging because of a lack of unique morphologic, immunophenotypic, molecular, and cytogenetic markers. The recurrent t(1;10)(p22;q24) translocation and/or TGFBR3/MGEA5 rearrangement was reported in 55 patients, with a relatively even distribution among HFLT, PHAT, and MIFS (17 HFLT, 15 MIFS, 13 MIFS/HFLT, and 10 PHAT). This indicates that current morphology-based diagnostic criteria do not identify reliably the subset of soft tissue tumor with t(1;10) translocation. Genetic heterogeneity of these tumors is supported by the recent detection of a mutually exclusive, second recurrent genetic change, t(7;17) TOM1L2-BRAF translocation or BRAF amplification, in a subset of MIFS.


Assuntos
Antígenos de Neoplasias/genética , Histona Acetiltransferases/genética , Hialuronoglucosaminidase/genética , Proteoglicanas/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Neoplasias de Tecidos Moles/genética , Neoplasias de Tecidos Moles/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Cromossomos Humanos Par 1/genética , Cromossomos Humanos Par 10/genética , Feminino , Rearranjo Gênico , Humanos , Masculino , Pessoa de Meia-Idade , Translocação Genética
15.
Int J Biol Macromol ; 121: 870-881, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30342141

RESUMO

Hyaluronic acid (HA) is a natural polymer with various molecular weights that specify multiple biological roles. Traditionally, HA is obtained from animal waste and conventional pathogenic streptococci. However, there are challenges in these processes such as the presence of exotoxins, hyaluronidase, and viral contamination. In order to reduce these problems, this study was conducted to produce HA using recombinant bacterium that is generally recognized as safe (GRAS), and thereafter increase production through experimental design. At first, some lactic acid bacteria were screened and evaluated for HA production. Accordingly, among the selected bacteria, Lactobacillus acidophilus PTCC1643 produced about 0.25 g HA/L in the 48th hour of cultivation, and was thus selected as an alternative host for heterologous HA production. An expression vector containing HA synthase genes was transformed into L. acidophilus by electroporation. Consequently, HA production increased to 0.4 g/L. Eventually, response surface method (RSM) was used, which increased HA production to 1.7 g/L. This is approximately 7-fold higher than that produced at first. The resulting HA was characterized by FTIR spectroscopy and its molecular weight was estimated using agarose gel electrophoresis. In conclusion, L. acidophilus could be a safe, effective, and novel HA producer with industrial potential and commercial prospects.


Assuntos
Meios de Cultura/química , Engenharia Genética , Ácido Hialurônico/biossíntese , Lactobacillus acidophilus/genética , Lactobacillus acidophilus/metabolismo , Eletroporação , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Lactobacillus acidophilus/crescimento & desenvolvimento , Peso Molecular
16.
Toxins (Basel) ; 10(12)2018 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-30567320

RESUMO

Venomous animals are found through a wide taxonomic range including cartilaginous fish such as the freshwater stingray Potamotrygon motoro occurring in South America, which can injure people and cause venom-related symptoms. Ensuring the efficacy of drug development to treat stingray injuries can be assisted by the knowledge of the venom composition. Here we performed a detailed transcriptomic characterization of the venom gland of the South American freshwater stingray Potamotrygon motoro. The transcripts retrieved showed 418 hits to venom components (comparably to 426 and 396 hits in other two Potamotrygon species), with high expression levels of hyaluronidase, cystatin and calglandulin along with hits uniquely found in P. motoro such as DELTA-alicitoxin-Pse1b, Augerpeptide hhe53 and PI-actitoxin-Aeq3a. We also identified undescribed molecules with extremely high expression values with sequence similarity to the SE-cephalotoxin and Rapunzel genes. Comparative analyses showed that despite being closely related, there may be significant variation among the venoms of freshwater stingrays, highlighting the importance of considering elicit care in handling different envenomation cases. Since hyaluronidase represents a major component of fish venom, we have performed phylogenetic and selective pressure analyses of this gene/protein across all fish with the available information. Results indicated an independent recruitment of the hyaluronidase into the stingray venom relative to that of venomous bony fish. The hyaluronidase residues were found to be mostly under negative selection, but 18 sites showed evidence of diversifying positive selection (P < 0.05). Our data provides new insight into stingray venom variation, composition, and selective pressure in hyaluronidase.


Assuntos
Proteínas de Peixes/genética , Venenos de Peixe/química , Hialuronoglucosaminidase/genética , /genética , Animais , Proteínas de Peixes/química , Transcriptoma
17.
J Mol Evol ; 86(9): 655-667, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30456442

RESUMO

Oviductal proteins play an important role in mammalian fertilization, as proteins from seminal fluid. However, in contrast with the latter, their phylogenetic evolution has been poorly studied. Our objective was to study in 16 mammals the evolution of 16 genes that encode oviductal proteins involved in at least one of the following steps: (1) sperm-oviduct interaction, (2) acrosome reaction, and/or (3) sperm-zona pellucida interaction. Most genes were present in all studied mammals. However, some genes were lost along the evolution of mammals and found as pseudogenes: annexin A5 (ANXA5) and deleted in malignant brain tumor 1 (DMBT1) in tarsier; oviductin (OVGP1) in megabat; and probably progestagen-associated endometrial protein (PAEP) in tarsier, mouse, rat, rabbit, dolphin, and megabat; prostaglandin D2 synthase (PTGDS) in microbat; and plasminogen (PLG) in megabat. Four genes [ANXA1, ANXA4, ANXA5, and heat shock 70 kDa protein 5 (HSPA5)] showed branch-site positive selection, whereas for seven genes [ANXA2, lactotransferrin (LTF), OVGP1, PLG, S100 calcium-binding protein A11 (S100A11), Sperm adhesion molecule 1 (SPAM1), and osteopontin (SPP1)] branch-site model and model-site positive selection were observed. These results strongly suggest that genes encoding oviductal proteins that are known to be important for gamete fertilization are subjected to positive selection during evolution, as numerous genes encoding proteins from mammalian seminal fluid. This suggests that such a rapid evolution may have as a consequence that two isolated populations become separate species more rapidly.


Assuntos
Tubas Uterinas/metabolismo , Tubas Uterinas/fisiologia , Mamíferos/genética , Acrossomo/fisiologia , Animais , Anexina A5/genética , Anexina A5/metabolismo , Evolução Biológica , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Evolução Molecular , Feminino , Glicoproteínas/genética , Humanos , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Lactoferrina/genética , Masculino , Filogenia , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Seleção Genética/genética , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo
18.
Int J Mol Sci ; 19(11)2018 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-30423867

RESUMO

Collagen peptide (CP) has beneficial effects on functions of the skin, such as skin barrier function and skin elasticity, in vivo. However, there are few studies investigating the mechanism underlying the potential effects of CP in skin epidermal moisturization after ultraviolet B (UVB) irradiation. In this study, we examined whether orally-administered CP affects the loss of skin hydration induced by UVB irradiation in hairless mice. SKH-1 hairless mice were orally administered CP at two doses (500 and 1000 mg/kg) for nine weeks, and the dorsal skin was exposed to UVB. The potential effects of CP were evaluated by measuring the transepidermal water loss (TEWL), skin hydration, wrinkle formation, and hyaluronic acid expression in the dorsal mice skin. We found that oral administration of CP increased skin hydration and decreased wrinkle formation compared to the UVB-irradiated group. Treatment of CP increased the mRNA and protein expression of hyaluronic acid synthases (HAS-1 and -2) concomitant with an increased hyaluronic acid production in skin tissue. The expression of hyaluronidase (HYAL-1 and 2) mRNA was downregulated in the CP-treated group. In addition, the protein expression of skin-hydrating factors, filaggrin and involucrin, was upregulated via oral administration of CP. In summary, these results show that oral administration of CP increases hyaluronic acid levels, which decreases during UVB photoaging. Therefore, we suggest that CP can be used as a nutricosmetic ingredient with potential effects on UVB-induced skin dehydration and moisture loss in addition to wrinkle formation.


Assuntos
Colágeno/administração & dosagem , Colágeno/uso terapêutico , Desidratação/tratamento farmacológico , Ácido Hialurônico/biossíntese , Peptídeos/administração & dosagem , Peptídeos/uso terapêutico , Pele/efeitos da radiação , Raios Ultravioleta , Administração Oral , Animais , Colágeno/farmacologia , Epiderme/efeitos dos fármacos , Epiderme/patologia , Epiderme/efeitos da radiação , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Masculino , Camundongos Pelados , Peptídeos/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Envelhecimento da Pele/efeitos dos fármacos , Envelhecimento da Pele/efeitos da radiação , Tilápia , Perda Insensível de Água
19.
J Proteome Res ; 17(11): 3941-3958, 2018 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-30270628

RESUMO

Snake venoms are complex mixtures mainly composed of proteins and small peptides. Crotoxin is one of the most studied components from Crotalus venoms, but many other components are less known due to their low abundance. The venome of Crotalus durissus terrificus, the most lethal Brazilian snake, was investigated by combining its venom gland transcriptome and proteome to create a holistic database of venom compounds unraveling novel toxins. We constructed a cDNA library from C. d. terrificus venom gland using the Illumina platform and investigated its venom proteome through high resolution liquid chromotography-tandem mass spectrometry. After integrating data from both data sets, more than 30 venom components classes were identified by the transcriptomic analysis and 15 of them were detected in the venom proteome. However, few of them (PLA2, SVMP, SVSP, and VEGF) were relatively abundant. Furthermore, only seven expressed transcripts contributed to ∼82% and ∼73% of the abundance in the transcriptome and proteome, respectively. Additionally, novel venom proteins are reported, and we highlight the importance of using different databases to perform the data integration and discuss the structure of the venom components-related transcripts identified. Concluding, this research paves the way for novel investigations and discovery of future pharmacological agents or targets in the antivenom therapy.


Assuntos
Venenos de Crotalídeos/química , Crotalus/fisiologia , Proteoma/isolamento & purificação , Transcriptoma , Sequência de Aminoácidos , Animais , Carboxipeptidases/genética , Carboxipeptidases/isolamento & purificação , Carboxipeptidases/metabolismo , Cromatografia Líquida/métodos , Fatores de Crescimento de Fibroblastos/genética , Fatores de Crescimento de Fibroblastos/isolamento & purificação , Fatores de Crescimento de Fibroblastos/metabolismo , Expressão Gênica , Biblioteca Gênica , Ontologia Genética , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/isolamento & purificação , Hialuronoglucosaminidase/metabolismo , Anotação de Sequência Molecular , Proteoma/genética , Proteoma/metabolismo , Alinhamento de Sequência , Análise de Sequência de RNA , Espectrometria de Massas em Tandem/métodos
20.
J Am Coll Cardiol ; 72(13): 1490-1503, 2018 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-30236312

RESUMO

BACKGROUND: Superficial erosion currently causes at least one-third of acute coronary syndromes (ACS), and its incidence is increasing. Yet, the underlying mechanisms in humans are still largely unknown. OBJECTIVES: The authors sought to assess the role of hyaluronan (HA) metabolism in ACS. METHODS: Peripheral blood mononuclear cells were collected from ACS (n = 66), stable angina (SA) (n = 55), and control (CTRL) patients (n = 45). The authors evaluated: 1) gene expression of hyaluronidase 2 (HYAL2) (enzyme degrading high-molecular-weight HA to its proinflammatory 20-kDa isoform) and of CD44v1, CD44v4, and CD44v6 splicing variants of HA receptor; and 2) HYAL2 and CD44 protein expression. Moreover, they compared HYAL2 and CD44 gene expression in ACS patients with plaque erosion (intact fibrous cap and thrombus) and in ACS patients with plaque rupture, identified by optical coherence tomography analysis. RESULTS: Gene expression of HYAL2, CD44v1, and CD44v6 were significantly higher in ACS as compared with SA (p = 0.003, p < 0.001, and p = 0.033, respectively) and CTRL subjects (p < 0.001, p < 0.001, and p = 0.009, respectively). HYAL2 protein expression was significantly higher in ACS than in SA (p = 0.017) and CTRL (p = 0.032), whereas no differences were found in CD44 protein expression. HYAL2 and CD44v6 gene expression was significantly higher in patients with plaque erosion than in those with plaque rupture (p = 0.015 and p = 0.029, respectively). CONCLUSIONS: HYAL2 and CD44v6 splicing variants seem to play an important role in ACS, in particular when associated with plaque erosion. After further validation, HYAL2 might represent a potentially useful biomarker for the noninvasive identification of this mechanism of coronary instability.


Assuntos
Síndrome Coronariana Aguda/metabolismo , Moléculas de Adesão Celular/genética , Hialuronoglucosaminidase/genética , Placa Aterosclerótica/diagnóstico por imagem , Síndrome Coronariana Aguda/genética , Idoso , Estudos de Casos e Controles , Moléculas de Adesão Celular/metabolismo , Feminino , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Expressão Gênica , Humanos , Receptores de Hialuronatos/genética , Receptores de Hialuronatos/metabolismo , Hialuronoglucosaminidase/metabolismo , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Processamento de Proteína , RNA Mensageiro/metabolismo , Tomografia de Coerência Óptica
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA