Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 181
J. bras. nefrol ; 41(3): 315-322, July-Sept. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1040245


Abstract Introduction: It is hypothesized that increased macrophage migration inhibitory factor (MIF) expression may contribute to diabetic nephropathy (DN) pathogenesis. The aim of the present study was to investigate the renal effects of MIF inhibition in a diabetic experimental model. Methods: Eighteen male Wistar rats (230 ± 20 g) were divided into three groups: 1) control, 2) diabetic (STZ, 50 mg/kg, dissolved in saline, ip), 3) diabetic + MIF antagonist (p425, 1 mg/kg per day, ip, on the 21th day, for 21 consecutive days). The treatment started since we founwd a significant increase in urine albumin excretion (UAE) rate in the diabetic rats in comparison with the control rats. The rats were kept individually in metabolic cages (8 AM-2 PM) and urine samples were collected in the 21 and 42th day. At the end, blood and tissue samples were collected for biochemical (BS, UPE, urine GAG, BUN, Cr, Na, and K) and histological analyses. Results: The results of this study showed that MIF antagonist (p425) significantly decreased urine protein and GAG excretion, urine protein/creatinine ratio, and serum BUN and Cr in the streptozotocin-induced DN in the rats. Pathological changes were significantly alleviated in the MIF antagonist (p425)-administered DN rats. Conclusion: Collectively, these data suggested that MIF antagonist (p425) was able to protect against functional and histopathological injury in the DN.

Resumo Introdução: Supõe-se que elevações da expressão do fator de inibição da migração de macrófagos (MIF) possam contribuir para a patogênese da nefropatia diabética (ND). O objetivo do presente estudo foi investigar os efeitos renais da inibição do MIF em um modelo experimental diabético. Métodos: Dezoito ratos Wistar machos (230 ± 20g) foram divididos em três grupos: 1) controle, 2) diabético (STZ 50 mg/kg dissolvida em soro fisiológico, IP), 3) diabético + antagonista do MIF (p425 1 mg/kg por dia IP no 21o dia por 21 dias consecutivos). O tratamento começou após a identificação de aumento significativo na albuminúria nos ratos diabéticos em relação aos controles. Os ratos foram mantidos individualmente em gaiolas metabólicas (8h-14h) e amostras de urina foram colhidas no 21o e no 42o dia. Ao final do estudo, amostras de sangue e tecido foram colhidas para análises bioquímicas (BS, excreção urinária de proteína, excreção urinária de GAGs, BUN, Cr, Na e K) e histológicas. Resultados: O presente estudo demonstrou que o antagonista do MIF (p425) diminuiu significativamente proteinúria, excreção urinária de GAGs , relação proteína/creatinina na urina, BUN e Cr no grupo com ND induzida por estreptozotocina. As alterações patológicas foram significativamente abrandadas nos ratos com ND que receberam antagonista do MIF (p425). Conclusão: Coletivamente, os dados sugerem que o antagonista do MIF (p425) teve efeito protetor contra lesões funcionais e histopatológicas da ND.

Animais , Masculino , Ratos , Fatores Inibidores da Migração de Macrófagos/antagonistas & inibidores , Oxirredutases Intramoleculares/antagonistas & inibidores , Substâncias Protetoras/uso terapêutico , Substâncias Protetoras/farmacologia , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/terapia , Glicemia , Ratos Wistar , Estreptozocina/farmacologia , Creatinina/urina , Creatinina/sangue , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/urina , Diabetes Mellitus Experimental/sangue , Nefropatias Diabéticas/urina , Nefropatias Diabéticas/patologia , Nefropatias Diabéticas/sangue , Albuminúria/tratamento farmacológico , Modelos Animais de Doenças , Glicosaminoglicanos/urina , Rim/patologia , Ativação de Macrófagos
An. bras. dermatol ; 94(3): 334-336, May-June 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1011112


Abstract: Cutaneous mucinoses are a complex and diverse group of connective tissue disorders characterized by the accumulation of mucin and/or glycosaminoglycan in the skin and adnexa. Cutaneous focal mucinosis appears as a solitary, asymptomatic, skin-colored to white papule, nodule, or plaque located anywhere on the body or in the oral cavity. It presents mainly in adults and is characterized on histopathology by mucin throughout the upper and mid dermis. We describe the dermoscopy of two cases of cutaneous focal mucinosis. Both lesions presented a nonspecific homogenous whitish pattern; the first case also exhibited a sharply demarcated yellow border.

Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Dermatopatias/patologia , Mucinoses/patologia , Dermoscopia , Glicosaminoglicanos , Mucinas
Braz. j. pharm. sci ; 52(4): 693-697, Oct.-Dec. 2016. tab, graf
Artigo em Inglês | LILACS | ID: biblio-951881


ABSTRACT Mucopolysaccharidoses (MPS) are a group of inherited metabolic disorders caused by deficiency of enzymes that degrade glycosaminoglycans (GAGs). Urinary excretion of GAGs is a common feature of MPS, and is considered their major biomarker. We aimed to adapt the GAG electrophoresis method to a commercial agarose gel which would be able to separate urinary GAGs in a simpler way with good sensitivity and reproducibility. Urine samples from patients previously diagnosed with MPS I, IV, and VI were used as electrophoretic standards. Samples from patients on enzyme replacement therapy (ERT) were also assessed. Commercial agarose gel electrophoresis was effective, showing proper definition and separation of GAG bands. Detection sensitivity exceeded 0.1 µg and band reproducibility were consistent. GAG bands quantified in urine samples from patients on ERT correlated very strongly (correlation coefficient = 0.98) with total GAG concentrations. This application of gel electrophoresis demonstrates the possibility of monitoring patients with MPS treated with ERT by analyzing separately the GAGs excreted in urine. We suggest this process should be applied to MPS screening as well as to follow-up of patients on treatment.

Humanos , Masculino , Feminino , Pré-Escolar , Mucopolissacaridoses/diagnóstico , Eletroforese em Gel de Ágar , Glicosaminoglicanos/uso terapêutico , Urina , Eletroforese/métodos
An. bras. dermatol ; 91(5): 595-600, Sept.-Oct. 2016. graf
Artigo em Inglês | LILACS | ID: biblio-827746


Abstract: Background: Heparanase is an enzyme that cleaves heparan sulfate chains. Oligosaccharides generated by heparanase induce tumor progression. Basal cell carcinoma and squamous cell carcinoma comprise types of nonmelanoma skin cancer. Objectives: Evaluate the glycosaminoglycans profile and expression of heparanase in two human cell lines established in culture, immortalized skin keratinocyte (HaCaT) and squamous cell carcinoma (A431) and also investigate the expression of heparanase in basal cell carcinoma, squamous cell carcinoma and eyelid skin of individuals not affected by the disease (control). Methods: Glycosaminoglycans were quantified by electrophoresis and indirect ELISA method. The heparanase expression was analyzed by quantitative RT-PCR (qRTPCR). Results: The A431 strain showed significant increase in the sulfated glycosaminoglycans, increased heparanase expression and decreased hyaluronic acid, comparing to the HaCaT lineage. The mRNA expression of heparanase was significantly higher in Basal cell carcinoma and squamous cell carcinoma compared with control skin samples. It was also observed increased heparanase expression in squamous cell carcinoma compared to the Basal cell carcinoma. Conclusion: The glycosaminoglycans profile, as well as heparanase expression are different between HaCaT and A431 cell lines. The increased expression of heparanase in Basal cell carcinoma and squamous cell carcinoma suggests that this enzyme could be a marker for the diagnosis of such types of non-melanoma cancers, and may be useful as a target molecule for future alternative treatment.

Humanos , Neoplasias Cutâneas/enzimologia , Carcinoma Basocelular/enzimologia , Carcinoma de Células Escamosas/enzimologia , Glucuronidase/metabolismo , Glicosaminoglicanos/metabolismo , RNA Mensageiro/metabolismo , Queratinócitos/metabolismo , Pálpebras/enzimologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Glucuronidase/genética , Glicosaminoglicanos/análise , Ácido Hialurônico/análise , Ácido Hialurônico/metabolismo
Acta ortop. bras ; 24(2): 98-101, Mar.-Apr. 2016. graf
Artigo em Inglês | LILACS | ID: lil-775079


Objetivo: Avaliar e comparar o comportamento dos glicosaminoglicanos(GAGs) na Doença de Dupuytren (DD). Métodos: Trata-se deum estudo experimental com 23 pacientes diagnosticados com DD.Tecidos coletados através de fasciectomia com incisão tipo Brunnerou McCash, foram avaliados por eletroforese para identificação dosGAGs. A quantificação foi realizada por imunofluorescência e dosagemdas proteínas para os diferentes tipos de glicosaminoglicanos.Os resultados coletados foram expressos em porcentagem e avaliadosestatisticamente. Resultados: Foi observado, na corrida eletroforética,aumento significativo dos GAGs em relação ao controle(p<0,05). Na imunofluorescência houve redução (23 vezes) do ácidohialurônico quando comparado ao controle (p<0,0001). Conclusão:Ficou evidenciado pelos resultados o aumento dos GAGs sulfatadosna doença de Dupuytren, principalmente do dermatam sulfato, egrande diminuição do ácido hialurônico na aponeurose palmar dosmesmos pacientes. Nível de Evidência III, Estudo Caso Controle.

Objective: To evaluate and compare the behavior of glycosaminoglycans(GAGs) in Dupuytren disease (DD). Methods: Thisis an experimental study with 23 patients diagnosed with DD.Tissue collected through fasciectomy with incision type Brunneror McCash were evaluated by electrophoresis for identificationof GAGs. The quantification was carried out by immunofluorescenceand dosage of proteins for different types of glycosaminoglycans.The results were expressed in percentage and statisticallyevaluated. Results: A significant increase was observed througheletrophoresis in GAGs, as compared to the control (p<0.05). Immunofluorescenceof hyaluronic acid was reduced (23 times) whencompared to the control (p<0.0001). Conclusion: An increase ofsulfated GAGs in Dupuytren’s disease, mainly dermatan sulfate,was evident from our results, as well as a pronounced decrease ofhyaluronic acid in the palmar aponeurosis from the same patients.Level of Evidence III, Case-Control Study.

Humanos , Contratura de Dupuytren , Eletroforese , Fáscia , Imunofluorescência , Glicosaminoglicanos , Mãos , Ácido Hialurônico , Proteoglicanas
Braz. j. med. biol. res ; 48(12): 1063-1070, Dec. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-762916


Damage to cartilage causes a loss of type II collagen (Col-II) and glycosaminoglycans (GAG). To restore the original cartilage architecture, cell factors that stimulate Col-II and GAG production are needed. Insulin-like growth factor I (IGF-I) and transcription factor SOX9are essential for the synthesis of cartilage matrix, chondrocyte proliferation, and phenotype maintenance. We evaluated the combined effect of IGF-I and SOX9 transgene expression on Col-II and GAG production by cultured human articular chondrocytes. Transient transfection and cotransfection were performed using two mammalian expression plasmids (pCMV-SPORT6), one for each transgene. At day 9 post-transfection, the chondrocytes that were over-expressing IGF-I/SOX9 showed 2-fold increased mRNA expression of the Col-II gene, as well as a 57% increase in Col-II protein, whereas type I collagen expression (Col-I) was decreased by 59.3% compared with controls. The production of GAG by these cells increased significantly compared with the controls at day 9 (3.3- vs 1.8-times, an increase of almost 83%). Thus, IGF-I/SOX9 cotransfected chondrocytes may be useful for cell-based articular cartilage therapies.

Humanos , Condrócitos/metabolismo , Colágeno Tipo II/biossíntese , Glicosaminoglicanos/biossíntese , Fator de Crescimento Insulin-Like I/metabolismo , Proteínas Matrilinas/biossíntese , Fatores de Transcrição SOX9/metabolismo , Transfecção/métodos , Cartilagem Articular/lesões , Cartilagem Articular/metabolismo , Colágeno Tipo II/análise , Matriz Extracelular/química , Expressão Gênica , Glicosaminoglicanos/análise , Fator de Crescimento Insulin-Like I/genética , Proteínas Matrilinas/genética , Cultura Primária de Células , Reação em Cadeia da Polimerase em Tempo Real , RNA Mensageiro/metabolismo , Fatores de Transcrição SOX9/genética , Espectrofotometria
Einstein (Säo Paulo) ; 13(4): 510-517, Oct.-Dec. 2015. tab, graf
Artigo em Português | LILACS | ID: lil-770491


ABSTRACT Objective To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Methods Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student’st test. Results The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). Conclusion The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues.

RESUMO Objetivo Determinar a presença de glicosaminoglicanos na matriz extracelular do tecido conjuntivo colorretal neoplásico e não neoplásico, tendo em vista seu papel central no desenvolvimento e na progressão dos tumores. Métodos Amostras de tecidos colorretais neoplásicos e não neoplásicos foram obtidas de 64 pacientes operados com carcinoma colorretal sem metástases a distância. As expressões de heparan sulfato, sulfato de condroitina e sulfato de dermatan e seus fragmentos foram analisadas por espectrometria de massa por ionização por electrospray, com técnica de extração e quantificação de glicosaminoglicanos após proteólise e eletroforese. Para análise estatística, utilizaram-se média, desvio padrão e teste t de Student. Resultados Em gel de agarose, os glicosaminoglicanos extraídos de tecido colorretal mostraram três bandas eletroforéticas. A espectrometria de massa por ionização por electrospray mostrou fragmentos de dissacarídeos característicos de glicosaminoglicanos e indicou sua característica estrutural. Alguns picos na espectrometria de massa por ionização por electrospray não foram caracterizados como fragmentos de açúcares, sugerindo a presença de fragmentos de proteínas estruturais dos proteoglicanos, formadas durante a purificação dos glicosaminoglicanos. A quantidade média de condroitina e dermatan aumentou no tecido neoplástico em relação ao tecido normal (p=0,01). Por outro lado, a quantidade média de heparan foi menor no tecido neoplásico em relação ao tecido normal (p=0,03). Conclusão O método empregado permitiu determinar o perfil estrutural dos glicosaminoglicanos nas amostras. Tecidos neoplásicos apresentaram maiores quantidades de sulfato de condroitina e sulfato de dermatan em comparação com os não neoplásicos, enquanto o sulfato de heparan foi encontrado em menores quantidades nos tecidos neoplásicos.

Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma/química , Neoplasias Colorretais/química , Matriz Extracelular/química , Glicômica/métodos , Glicosaminoglicanos/análise , Carcinoma/patologia , Sulfatos de Condroitina/análise , Neoplasias Colorretais/patologia , Tecido Conjuntivo/química , Progressão da Doença , Dermatan Sulfato/análise , Eletroforese em Gel de Poliacrilamida , Heparitina Sulfato/análise , Membrana Mucosa/metabolismo , Proteólise , Espectrometria de Massas por Ionização por Electrospray
Acta sci., Biol. sci ; 37(4): 411-417, Oct.-Dec. 2015. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-876365


The production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920 with varying rates of pH (6.0, 7.0, 8.0), temperature (34; 37; 40°C), agitation (100, 150, 200 rpm), glucose (10, 20, 30 g L -1) and yeast extract concentration (10, 20, 30 g L -1) was evaluated by statistical approaches. The best conditions for the production of hyaluronic acid was pH 8.0, 37°C and 100 rpm in a medium containing 30 g L- 1 glucose and yeast extract, for a production of 0.787 g L- 1. Temperature, pH and yeast extract were significant variables (p < 0.05). Yeast extract and pH had a positive effect on the production of the polymer. Lactate, formate and acetate synthesis were also analyzed. Current assay showed the feasibility of statistical tools to optimize the physical and nutritional parameters for the production of hyaluronic acid and the improvement of the fermentation process.

A produção de ácido hialurônico por Streptococcus zooepidemicus ATCC 39920 foi avaliada variando pH (6,0; 7,0, 8,0), temperatura (34; 37; 40°C), agitação (100, 150, 200 rpm) e concentração de glicose (10, 20, 30 g L-1) e extrato de levedura (10, 20, 30 g L-1) por metodologias estatísticas. A condição otimizada foi pH 8,0, 37°C e 100 rpm, em meio contendo 30 g L-1 de glicose e extrato de levedura atingindo a produção de 0,787 g L-1. O pH, temperatura e extrato de levedura foram as variáveis significativas (p < 0,05). Extrato de levedura e pH apresentaram efeito positivo para a produção do polímero. A síntese de ácido lático, fórmico e acético também foi analisada. Este estudo demonstra a viabilidade de utilização de ferramentas estatísticas para otimizar os parâmetros físicos e nutricionais para a produção de ácido hialurônico, permitindo a melhoria do processo fermentativo.

Glicosaminoglicanos , Ácido Hialurônico , Interações Microbianas , Fenômenos Fisiológicos da Nutrição , Fatores Físicos e Químicos
São Paulo; s.n; 2015. [101] p. ilus, tab, graf.
Tese em Português | LILACS | ID: biblio-871516


INTRODUÇÃO: A matriz extracelular no endométrio fornece vasta gama de sinais envolvidos em diferentes processos celulares, tais como morte celular e proliferação. Neste sentido, os glicosaminoglicanos e os proteoglicanos, juntamente com os fatores de crescimento, modulam várias etapas da angiogênese, proliferação celular e remodelação do estroma, o que pode ser importante para o fluxo menstrual regular e a redução dos processos proliferativos. Além disso, são importantes para a adequada interação maternofetal. OBJETIVO: Avaliar a concentração de ácido hialurônico, das enzimas de biossíntese do ácido hialurônico - hialurônico sintases (HAS1, HAS2 e HAS3) e dos pequenos proteoglicanos ricos em leucina (decorim, biglicam, lumicam e fibromodulina) no endométrio de pacientes com síndrome dos ovários policísticos (SOP) e de mulheres eumenorreicas. MÉTODOS: Foram analisadas 20 amostras de endométrio, 10 provenientes de pacientes com SOP e 10 de mulheres com ciclos menstruais regulares na fase proliferativa do ciclo, com idade variando entre 20 e 35 anos, atendidas na Divisão de Clínica Ginecológica do Hospital das Clínicas da FMUSP (HC-FMUSP). A determinação do perfil e da concentração do ácido hialurônico foi efetuada por método bioquímico de ensaio fluorimétrico (ELISA-like). A sua localização no tecido endometrial, assim como as dosagens das enzimas sintases (HAS1, HAS2 e HAS3) e dos pequenos proteoglicanos ricos em leucina (decorim, biglicam, lumicam e fibromodulina e), foi feita por imunoistoquímica e "western blotting". Para a análise dos resultados foi utilizado o teste t de student (p<=0,05). Os cálculos foram realizados com auxílio do programa SPSS versão13 (SPSS, Chicago, IL). RESUTADOS: Houve maior concentração de ácido hialurônico no endométrio de mulheres eumenorreicas na fase proliferativa do ciclo menstrual do que no das com síndrome dos ovários policísticos. Com relação às sintases do ácido hialurônico, observou-se maior reatividade de HAS1 e HAS2 e ...

INTRODUCTION: Endometrium extracellular matrix provides wide range of signals at different cellular levels like cell death and proliferation. In this regard, glycosaminoglycans and proteoglycans, along with growth factors, modulate various stages of angiogenesis, cell proliferation and remodeling of the stroma, which can be important for regulating menses and reducing the proliferative processes. Additionally, it is important for proper fetal-maternal interactions. OBJECTIVE: Evaluate hyaluronic acid concentration, the enzymes of hyaluronic acid synthases (HAS1, HAS2 and HAS3) and small leucine-rich proteoglycans (decorin, lumican, fibromodulim and biglycan) in the endometrium of patients with polycystic ovary syndrome (PCOS) and eumenorrheic women. METHODS: A total of 20 endometrial samples from 10 patients with PCOS and 10 women with regular menstrual cycles in the proliferative phase, with ages ranging between 20 and 35 years, attended at Gynecology Division of Clinical Hospital of the FMUSP (HC-USP). Profile determination and the concentration of hyaluronic acid were performed by biochemical method of fluorimetric assay (ELISA-like). Its location in the endometrial tissue as well as the dosage of enzymes synthases (HAS1, HAS2 and HAS3) and small leucine-rich proteoglycans (decorin, lumican, fibromodulim and biglycan) was done by immunohistochemistry and western blotting. To analyze the results Student t test was used (p < 0.05). The calculations were performed with software SPSS version 13. RESULTS: A higher concentration of hyaluronic acid in eumenorrheic women endometrium in proliferative phase when compared with polycystic ovary syndrome. Regarding hyaluronic acid synthases, there was a higher HAS1 and HAS2 reactivity and lower HAS3 reactivity in the PCOS endometrium compared to women with regular menstrual cycles in the proliferative phase. Decorin and lumican showed higher immunoreactivity in PCOS endometrium. CONCLUSIONS: PCOS patients have a lower...

Humanos , Feminino , Adulto Jovem , Adulto , Endométrio , Glicosaminoglicanos , Ácido Hialurônico , Síndrome do Ovário Policístico , Proteoglicanas
Invest. clín ; 55(4): 365-370, dic. 2014. ilus
Artigo em Inglês | LILACS | ID: lil-783090


Mucopolysaccharidoses are a group of lysosomal storage disorders caused by deficiency of enzymes catalyzing the degradation of glycosaminoglycans. Mucopoly-saccharidosis I can present a wide range of phenotypic characteristics with three major recognized clinical entities: Hurler and Scheie syndromes represent phenotypes at the severe and mild ends of the clinical spectrum, respectively, and the Hurler-Scheie syndrome is intermediate in phenotypic expression. These are caused by the deficiency or absence of α-L-iduronidase, essential to the metabolism of both dermatan and heparan sulfate, and it is encoded by the IDUA gene. We report the case of a 34-year-old male patient with enzymatic deficiency of α-L-iduronidase, accumulation of its substrate and a previously unreported mutation in the IDUA gene that developed a phenotype of Scheie syndrome.

Las mucopolisacaridosis son un grupo de trastornos de almacenamiento lisosomal causada por la deficiencia de enzimas que catalizan la degradación de glicosaminoglicanos. La mucopolisacaridosis tipo I puede presentar un amplio rango de características fenotípicas englobadas en tres entidades clínicas reconocidas: los síndromes de Hurler y Scheie representan los fenotipos graves y leves del espectro clínico, respectivamente y el síndrome de Hurler-Scheie intermedio en la expresión fenotípica. Estos son causados por la deficiencia o ausencia de la α-L-iduronidasa esencial para el metabolismo del dermatán y el heparán sulfato y es codificada por el gen IDUA. Se presenta el caso de paciente masculino de 34 años de edad con deficiencia enzimática de α-L-iduronidasa, acumulación de su sustrato y una mutación en el gen IDUA, no reportada previamente, que desarrolló un fenotipo del síndrome de Scheie.

Adulto , Humanos , Masculino , Iduronidase/genética , Mutação de Sentido Incorreto , Mucopolissacaridose I/genética , Mutação Puntual , Substituição de Aminoácidos , Progressão da Doença , Dermatan Sulfato/urina , Éxons/genética , Glicosaminoglicanos/metabolismo , Heterozigoto , Deformidades Adquiridas da Mão/genética , Íntrons/genética , Imagem por Ressonância Magnética , Mucopolissacaridose I/urina , Fenótipo , Deleção de Sequência , Avaliação de Sintomas , Compressão da Medula Espinal/etiologia , Compressão da Medula Espinal/patologia
Arq. gastroenterol ; 51(4): 309-315, Oct-Dec/2014. graf
Artigo em Inglês | LILACS | ID: lil-732204


Context Cholestasis produces hepatocellular injury, leukocyte infiltration, ductular cells proliferation and fibrosis of liver parenchyma by extracellular matrix replacement. Objective Analyze bile duct ligation effect upon glycosaminoglycans content and matrix metalloproteinase (MMPs) activities. Methods Animals (6-8 weeks; n = 40) were euthanized 2, 7 or 14 days after bile duct ligation or Sham-surgery. Disease evolution was analyzed by body and liver weight, seric direct bilirubin, globulins, gamma glutamyl transpeptidase (GGT), alkaline phosphatase (Alk-P), alanine and aspartate aminotransferases (ALT and AST), tissue myeloperoxidase and MMP-9, pro MMP-2 and MMP-2 activities, histopathology and glycosaminoglycans content. Results Cholestasis caused cellular damage with elevation of globulins, GGT, Alk-P, ALT, AST. There was neutrophil infiltration observed by the increasing of myeloperoxidase activity on 7 (P = 0.0064) and 14 (P = 0.0002) groups which leads to the magnification of tissue injuries. Bile duct ligation increased pro-MMP-2 (P = 0.0667), MMP-2 (P = 0.0003) and MMP-9 (P<0.0001) activities on 14 days indicating matrix remodeling and establishment of inflammatory process. Bile duct ligation animals showed an increasing on dermatan sulfate and/or heparan sulfate content reflecting extracellular matrix production and growing mitosis due to parenchyma depletion. Conclusions Cholestasis led to many changes on rats’ liver parenchyma, as so as on its extracellular matrix, with major alterations on MMPs activities and glycosaminoglycans content. .

Contexto Colestase produz lesão hepatocelular, infiltração leucocitária, proliferação de células ductulares e fibrose do parênquima hepático por matriz extracelular. Objetivo Analisar os efeitos da ligação do ducto biliar sobre conteúdo de glicosaminoglicanos e atividade de metaloproteinases de matriz (MMP). Métodos Animais (6-8 semanas; n = 40) foram eutanasiados 2, 7 ou 14 dias após ligação do ducto biliar ou falsa ligação. A evolução da doença foi analisada por peso corporal e do fígado, concentrações séricas de bilirrubina direta, globulinas, gama glutamil transpeptidase (GGT), fosfatase alcalina (Alk-P), alanina e aspartato aminotransfesases (ALT e AST), alterações teciduais de mieloperoxidase e metaloproteinases (MMP-9, pro MMP-2 e MMP-2), histopatologia e conteúdo de glicosaminoglicanos. Resultados A colestase causou dano celular com elevação dos níveis séricos de globulinas, GGT, Alk-P, ALT e AST. Houve também infiltração leucocitária observada pelo aumento na atividade de mieloperoxidase nos grupos 7 (P = 0,0064) e 14 dias (P = 0,0002) o que leva ao aumento das lesões no tecido. Ligação do ducto biliar aumentou as atividades de pro MMP-2 (P = 0,0677), MMP-2 (P = 0,0003) e MMP-9 (P<0,0001) aos 14 dias indicando remodelamento da matriz e estabelecimento de processo inflamatório. Animais com ligação do ducto biliar mostraram um aumento do conteúdo de dermatam sulfato e/ou heparam sulfato refletindo a produção de matriz extracelular e aumento de mitose devido a depleção do parênquima hepático. Conclusões Colestase causou várias mudanças no parênquima hepático de ratos, bem como em sua matriz extracelular, com importantes alterações na atividade ...

Animais , Masculino , Colestase Extra-Hepática/metabolismo , Matriz Extracelular/química , Glicosaminoglicanos/metabolismo , Metaloproteases/metabolismo , Glicosaminoglicanos/análise , Metaloproteases/análise , Ratos Wistar
Braz. j. med. biol. res ; 47(8): 637-645, 08/2014. tab, graf
Artigo em Inglês | LILACS | ID: lil-716279


Tissue engineering encapsulated cells such as chondrocytes in the carrier matrix have been widely used to repair cartilage defects. However, chondrocyte phenotype is easily lost when chondrocytes are expanded in vitro by a process defined as “dedifferentiation”. To ensure successful therapy, an effective pro-chondrogenic agent is necessary to overcome the obstacle of limited cell numbers in the restoration process, and dedifferentiation is a prerequisite. Gallic acid (GA) has been used in the treatment of arthritis, but its biocompatibility is inferior to that of other compounds. In this study, we modified GA by incorporating sulfamonomethoxine sodium and synthesized a sulfonamido-based gallate, JJYMD-C, and evaluated its effect on chondrocyte metabolism. Our results showed that JJYMD-C could effectively increase the levels of the collagen II, Sox9, and aggrecan genes, promote chondrocyte growth, and enhance secretion and synthesis of cartilage extracellular matrix. On the other hand, expression of the collagen I gene was effectively down-regulated, demonstrating inhibition of chondrocyte dedifferentiation by JJYMD-C. Hypertrophy, as a characteristic of chondrocyte ossification, was undetectable in the JJYMD-C groups. We used JJYMD-C at doses of 0.125, 0.25, and 0.5 µg/mL, and the strongest response was observed with 0.25 µg/mL. This study provides a basis for further studies on a novel agent in the treatment of articular cartilage defects.

Animais , Coelhos , Benzamidas/síntese química , Desdiferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Fenótipo , Pirimidinas/síntese química , Agrecanas/genética , Agrecanas/metabolismo , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Benzamidas/farmacologia , Sobrevivência Celular , Desdiferenciação Celular/imunologia , Condrócitos/citologia , Condrócitos/metabolismo , Condrogênese/efeitos dos fármacos , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Glicosaminoglicanos/análise , Imuno-Histoquímica , Citometria de Varredura a Laser , Cultura Primária de Células , Pirimidinas/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Engenharia Tecidual
Arch. cardiol. Méx ; 84(2): 110-116, abr.-jun. 2014. ilus
Artigo em Espanhol | LILACS | ID: lil-732015


El glucocáliz endotelial es una capa constituida por glucosaminoglicanos, proteoglicanos y glucoproteínas que cubre al endotelio en su cara luminal. La participación del deterioro del glucocáliz endotelial parece esencial en los pasos iniciales de la fisiopatología de la aterosclerosis, de las complicaciones microangiopáticas de la diabetes mellitus y de la enfermedad venosa crónica. Los factores de riesgo de la aterosclerosis como la hipercolesterolemia, la hiperglucemia, la inflamación, el exceso de sodio y las fuerzas de tensión alteradas causan deterioro del glucocáliz. Esto provoca disfunción endotelial y permite la filtración de lipoproteínas (LDL) y de leucocitos al espacio subendotelial, iniciando la formación de la placa de ateroma. En la diabetes el glucocáliz adelgazado, principalmente por estrés oxidativo, posibilita la filtración de proteínas (albuminuria) y el trastorno endotelial de la microangiopatía. La hipertensión venosa crónica altera las fuerzas de tensión y daña el glucocáliz, lo que permite la filtración de leucocitos a las partes más profundas de la pared venosa, iniciando la inflamación y el deterioro morfológico y funcional de las venas que lleva a la enfermedad venosa crónica. El tratamiento con glucosaminoglicanos (sulodexida) logra prevenir o revertir el daño al glucocáliz endotelial y algunas de sus consecuencias; es eficaz en la enfermedad venosa crónica, especialmente con úlceras venosas. También ha sido útil en aterosclerosis obliterante de miembros inferiores y en la nefropatía diabética con albuminuria.

Endothelial glycocalyx is a layer composed by glycosaminoglycans, proteoglycans and glycoproteins attached to the vascular endothelial luminal surface. Shredding of glycocalyx appears as an essential initial step in the pathophysiology of atherosclerosis and microangiopathic complications of diabetes mellitus, as well as in chronic venous disease. Atherosclerosis risk factors, as hypercholesterolemia (LDL), hyperglycemia, inflammation, salt excess and altered shear stress can damage glycocalyx. This lead to endothelial dysfunction and allows LDL and leukocytes to filtrate to the subendothelial space initiating atheroma plaque formation. Degradation of glycocalyx in diabetes mellitus is mainly due to oxidative stress and enables protein filtration (albuminuria) and endothelial disorder of microangiopathy. Chronic venous hypertension brings to altered shears stress which results in shredded glycocalyx, this allows leukocytes to migrate into venous wall and initiate inflammation leading to morphologic and functional venous changes of the chronic venous disease. Treatment with glycosaminoglycans (sulodexide) prevents or recovers the damaged glycocalyx and several of its consequences. This drug improves chronic venous disease and promotes healing of chronic venous ulcers. It has also been useful in peripheral arterial obstructive disease and in diabetic nephropathy with albuminuria.

Humanos , Angiopatias Diabéticas/etiologia , Endotélio Vascular , Glicocálix/fisiologia , Doenças Vasculares/etiologia , Aterosclerose/etiologia , Aterosclerose/patologia , Doença Crônica , Angiopatias Diabéticas/tratamento farmacológico , Angiopatias Diabéticas/patologia , Endotélio Vascular/química , Glicocálix/química , Glicocálix/efeitos dos fármacos , Glicosaminoglicanos/uso terapêutico , Doenças Vasculares/tratamento farmacológico , Doenças Vasculares/patologia , Pressão Venosa/fisiologia
Int. braz. j. urol ; 40(1): 72-79, Jan-Feb/2014. tab, graf
Artigo em Inglês | LILACS | ID: lil-704176


Introduction: Painful bladder syndrome/interstitial cystitis (PBS/IC) pathogenesis is not fully known, but evidence shows that glycosaminoglycans (GAG) of bladder urothelium can participate in its genesis. The loss of these compounds facilitates the contact of urine compounds with deeper portions of bladder wall triggering an inflammatory process. We investigated GAG in urine and tissue of PBS/IC and pure stress urinary incontinence (SUI) patients to better understand its metabolism. Materials and Methods: Tissue and urine of 11 patients with PBS/IC according to NIDDK criteria were compared to 11 SUI patients. Tissue samples were analyzed by histological, immunohistochemistry and immunofluorescence methods. Statistical analysis were performed using t Student test and Anova, considering significant when p < 0.05. Results: PBS/IC patients had lower concentration of GAG in urine when compared to SUI (respectively 0.45 ± 0.11 x 0.62 ± 0.13 mg/mg creatinine, p < 0.05). However, there was no reduction of the content of GAG in the urothelium of both groups. Immunofluorescence showed that PBS/IC patients had a stronger staining of TGF-beta, decorin (a proteoglycan of chondroitin/dermatan sulfate), fibronectin and hyaluronic acid. Conclusion: the results suggest that GAG may be related to the ongoing process of inflammation and remodeling of the dysfunctional urothelium that is present in the PBS/IC. .

Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Cistite Intersticial/metabolismo , Glicosaminoglicanos/metabolismo , Incontinência Urinária por Estresse/metabolismo , Biópsia , Creatinina/urina , Cistite Intersticial/patologia , Imunofluorescência , Glicosaminoglicanos/análise , Ácido Hialurônico/urina , Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo Real , Bexiga Urinária/patologia , Incontinência Urinária por Estresse/patologia , Urotélio/metabolismo , Urotélio/patologia
Rev. chil. obstet. ginecol ; 79(4): 288-293, 2014. ilus
Artigo em Espanhol | LILACS | ID: lil-724829


Los mecanismos fisiopatológicos de la malaria placentaria son hasta el momento poco comprendidos, y el daño placentario derivado de la infección por Plasmodium spp se ha relacionado con eventos adversos del embarazo que afectan directamente el desarrollo del feto. Las concentraciones placentarias de algunas citocinas como la IL-10, TNF-alfa y TGF-beta y glicosaminoglicanos como el CSA, HA y HS podrían estar participando de forma reguladora en los eventos inflamatorios placentarios durante la infección por Plasmodium spp.

The pathophysiological mechanisms of placental malaria are until now poorly understood and the placental damage resulting from infection by Plasmodium spp has been linked to adverse pregnancy events that directly affect fetal development. Placental concentrations of some cytokines such as IL-10, TNF-alpha and TGF-beta and glycosaminoglycans such as CSA, HA and HS could be involved in a regulatory role in placental inflammation during infection by Plasmodium spp.

Humanos , Feminino , Gravidez , Glicosaminoglicanos , Malária/imunologia , Malária/parasitologia , Placenta/imunologia , Placenta/parasitologia , Plasmodium , Complicações Parasitárias na Gravidez
Int. j. morphol ; 31(4): 1331-1339, Dec. 2013. ilus
Artigo em Inglês | LILACS | ID: lil-702314


The ontogenesis of the gastroesophageal mucosa involves morphological alterations related to its structure and the function of each segment. The present study describes the histogenesis of the mucus-secreting epithelium and glands of the esophagus, gizzard, and proventriculus of the chicken (Gallus gallus), and identifies alterations in the secretion pattern of glycosaminoglycans (GAG's). We analyzed 38 chicken embryos, processed the material collected following the histological routine, and then stained it with hematoxylin-eosin for the analysis of tissue structure and with Gomori's trichrome for the identification of conjunctive tissue and collagen fibers. We used the PAS histochemical technique for the analysis of neutral GAG's and the AB pH 2.5 histochemical technique for the analysis of acid GAG's. The embryos at late stage of development had the esophagus wall composed of four layers: mucosa, submucosa, muscularis, and serosa, whereas the proventriculus and the gizzard were composed of three layers: mucosa, muscularis, and serosa. In all three segments, we identified the superficial epithelium as mucus-secreting; in the esophagus this epithelium was mucus-secreting only at the initial development stages. The proventricular glands began to form at the initial development stages, whereas the tubular glands began to form in the gizzard just after the 15th day. The differences in the production of GAG's in these regions of the digestive tract are related to development stages, functions, and physiological requirements of each segment, and to the gradual adjustment of the body to the post-hatching life.

La morfogénesis de la mucosa gastroesofágica implica alteraciones morfológicas relacionadas con su estructura y la función de cada segmento. El presente estudio describe la histogénesis del epitelio secretor mucinoso y de las glándulas del esófago, molleja y proventrículo del pollo (G. gallus), ademas de identificar los cambios em el patrón de secreción de glicocosaminoglicanos (GAG's). Se utilizaron treinta y ocho embriones de pollo. El material recogido fue procesado de acuerdo a la rutina histológica y posteriormente las secciones se tiñieron con hematoxilina-eosina para su análisis histológico, con tricrómico de Gomori para identificar el tejido conectivo y las fibras de colágeno y con PAS y AB pH 2,5 para el análisis GAG's neutro y ácido. En una etapa avanzada de desarrollo de los embriones, se pudo obervar en la pared del esófago cuatro capas: mucosa, submucosa, muscular y serosa, mientras que el proventrículo y molleja se muestra constituida por tres capas: mucosa, muscular y serosa. En los tres segmentos de la superficie, el epitelio se identificó como mucinoso y en el esófago sólo en la etapa inicial de desarrollo. Las glándulas del proventrículo se empiezan a formar en las primeras etapas de desarrollo, mientras que en las glándulas tubulares de la molleja comienzan su sólo después del día 15. Las diferencias en la producción de GAG's en estas regiones del tracto digestivo están relacionadas con las etapas de desarrollo, las funciones y necesidades fisiológicas de cada segmento del cuerpo y se adapta gradualmente a la vida después de la eclosión.

Animais , Mucosa Gástrica/anatomia & histologia , Mucosa Gástrica/crescimento & desenvolvimento , Mucosa Gástrica , Galinhas/anatomia & histologia , Embrião de Galinha , Glicosaminoglicanos , Morfogênese
Braz. j. med. biol. res ; 46(7): 567-573, ago. 2013. tab, graf
Artigo em Inglês | LILACS | ID: lil-682402


Glycosaminoglycans (GAGs) participate in a variety of processes in the kidney, and evidence suggests that gender-related hormones participate in renal function. The aim of this study was to analyze the relationship of GAGs, gender, and proteinuria in male and female rats with chronic renal failure (CRF). GAGs were analyzed in total kidney tissue and 24-h urine of castrated (c), male (M), and female (F) Wistar control (C) rats (CM, CMc, CF, CFc) and after 30 days of CRF induced by 5/6 nephrectomy (CRFM, CRFMc, CRFF, CRFFc). Total GAG quantification and composition were determined using agarose and polyacrylamide gel electrophoresis, respectively. Renal GAGs were higher in CF compared to CM. CRFM presented an increase in renal GAGs, heparan sulfate (HS), and proteinuria, while castration reduced these parameters. However, CRFF and CRFFc groups showed a decrease in renal GAGs concomitant with an increase in proteinuria. Our results suggest that, in CRFM, sex hormones quantitatively alter GAGs, mainly HS, and possibly the glomerular filtration barrier, leading to proteinuria. The lack of this response in CRFMc, where HS did not increase, corroborates this theory. This pattern was not observed in females. Further studies of CRF are needed to clarify gender-dependent differences in HS synthesis.

Animais , Feminino , Masculino , Castração , Glicosaminoglicanos/urina , Hormônios Esteroides Gonadais/deficiência , Falência Renal Crônica/metabolismo , Rim/química , Proteinúria/urina , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Taxa de Filtração Glomerular , Glicosaminoglicanos/isolamento & purificação , Heparitina Sulfato/urina , Falência Renal Crônica/cirurgia , Rim/cirurgia , Nefrectomia , Distribuição Aleatória , Ratos Wistar , Fatores Sexuais
Neumol. pediátr ; 8(1): 27-33, 2013. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-701687


Mucopolysaccharidosis (MPS) are part of the so-called lysosomal diseases, in which the deposit of different glycosaminoglycans, depending on the enzyme deficit, generates multi-systemic manifestations, being the respiratory system one of the most affected and associated with significant morbidity and mortality. Different types of MPS show a variable degree of organ compromise even from the early stages of life: obstruction of the upper airway of varying degree, persistent rhinorrhea, otitis media, obstructive pathology of the peripheral airway, pneumonias or other infections associated with a poor mucociliary drainage are the main manifestations presented by patients. The compromise of the neurological and musculoskeletal system also brings the compromise of the respiratory pump. From that perspective the approach must be multidisciplinary, since there are several organs and systems involved. Current therapy is directed to replace the deficient enzyme but it’s available only for some of them, which delays the progression of the disease but does not stop it, even more so there is no effect on the central nervous system, being the cognitive compromise inevitable. Bone marrow transplant is a therapy not exempt of complications, but capable of changing the progression of the disease in its early stages. Therapeutic approach is based on support measures and treatment of concurrent complications, both of which will be discussed in the following article.

Las Mucopoliscaridosis (MPS) son parte de las denominadas enfermedades lisosomales. El depósito de los distintos glicosaminoglicanos comprometidos, dependiendo del déficit enzimático, genera manifestaciones multisistémicas, en donde el sistema respiratorio es uno de los principales afectados y que se asocia con morbilidad y mortalidad significativa. Los diferentes tipos de MPS presentan un grado variable de compromiso desde etapas precoces de la vida, síntomas de obstrucción de vía aérea superior de grado variable, rinorrea persistente, otitis media, patología obstructiva de vía aérea periférica, neumonías o infecciones asociadas a un mal drenaje mucociliar son las principales manifestaciones que los pacientes presentan. El compromiso neurológico y musculo esquelético, trae consigo además el compromiso de la bomba respiratoria. Desde esa perspectiva el enfoque debe ser multidisciplinario, ya que el compromiso abarca varios órganos y sistemas. Las actuales terapias están dirigidas a reemplazar la enzima deficitaria, disponibles sólo para algunas de ellas, esto trae consigo el retardo de la evolución de la enfermedad pero no lo evita, considerando que más aun no tiene ningún efecto sobre el sistema nervioso central, por lo que el compromiso cognitivo es inevitable. El trasplante de médula es una terapia no exenta de complicaciones, pero que es capaz de cambiar la progresión de la enfermedad en las etapas precoces de ella. El enfoque terapéutico se basa en terapia de sostén y el manejo de las distintas complicaciones que se van dando, siendo éstos los ejes del siguiente artículo.

Humanos , Criança , Doenças Respiratórias/etiologia , Mucopolissacaridoses/complicações , Apneia Obstrutiva do Sono/etiologia , Doenças Respiratórias , Glicosaminoglicanos , Mucopolissacaridoses/classificação , Mucopolissacaridoses , Obstrução das Vias Respiratórias/etiologia
Clinics ; 67(8): 939-944, Aug. 2012. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-647799


OBJECTIVES: The promotion of extracellular matrix synthesis by chondrocytes is a requisite part of an effective cartilage tissue engineering strategy. The aim of this in vitro study was to determine the effect of bi-axial cyclic mechanical loading on cell proliferation and the synthesis of glycosaminoglycans by chondrocytes in threedimensional cultures. METHOD: A strain comprising 10% direct compression and 1% compressive shear was applied to bovine chondrocytes seeded in an agarose gel during two 12-hour conditioning periods separated by a 12-hour resting period. RESULTS: The bi-axial-loaded chondrocytes demonstrated a significant increase in glycosaminoglycan synthesis compared with samples exposed to uni-axial or no loading over the same period (p<0.05). The use of a free-swelling recovery period prior to the loading regime resulted in additional glycosaminoglycan production and a significant increase in DNA content (p<0.05), indicating cell proliferation. CONCLUSIONS: These results demonstrate that the use of a bi-axial loading regime results in increased matrix production compared with uni-axial loading.

Animais , Bovinos , Condrócitos/metabolismo , Matriz Extracelular/metabolismo , Glicosaminoglicanos/biossíntese , Regulação para Cima/fisiologia , Proliferação de Células , Células Cultivadas , Força Compressiva , Condrócitos/citologia , Matriz Extracelular/genética , Sefarose , Estresse Mecânico , Fatores de Tempo , Engenharia Tecidual/métodos