Navegando por Palavras-chave "Glycosaminoglycans"

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    Ação de um Peptídeo Derivado do Lopap em Cultura de Fibroblastos e em Modelo Experimental de Lesão Cutânea em Ratos
    (Universidade Federal de São Paulo (UNIFESP), 2009-06-24) Wlian, Luana [UNIFESP]; Tavassi, Ana Marisa Chudzinski [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    The process of tissue repair is a complex of reactions between cells and biochemical mediators that are triggered from an injury. This study evaluated the effect of peptide 4 derived from the amino acid sequence of Lopap (Lonomia obliqua prothrombin activator protease) in culture of fibroblasts to evaluate the expression of molecules of extracellular matrix and the membrane receptors of interleukin-8 and in experimental model of skin lesion in rats to examine the effect of the molecule in the process of tissue repair. We used 40 male Wistar rats with 6 to 8 weeks, randomized to 5 experimental groups. After anesthesia, the rats were submitted to surgery, where eight circular full-thickness of skin from 6 mm of diameter were surrounded in the back and treated with saline or with peptide 4. After different periods (days 3, 7, 14 and 21 days after wounding) were carried out tests to evaluate the process of tissue repair between the groups. We conclude that peptide 4 is able to stimulate fibroblasts in culture and promote the acceleration of the process of tissue repair in vivo model with greater deposition of collagen, glycosaminoglycans and better tissue reorganization.
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    Ação dos esteroides ovarianos sobre a quantidade dos glicosaminoglicanos sulfatados e do ácido hialurônico nas mamas de ratas ovariectomizadas
    (Universidade Federal de São Paulo (UNIFESP), 2016-11-17) Torres, Sueli Maria Preda dos Santos [UNIFESP]; Simoes, Manuel de Jesus [UNIFESP]; http://lattes.cnpq.br/5987164343458678; http://lattes.cnpq.br/2539410198240163; Universidade Federal de São Paulo (UNIFESP)
    The extracellular matrix (ECM) act an essential role in the function of mammary epithelial cells and glycosaminoglycans have long been attracting clinical interest in all areas of medicine for their important role in cell recognition, migration, cell proliferation and differentiation. OBJECTIVE: Evaluate the morphology and the concentration of glycosaminoglycans in the breast of female rats treated with estrogen and/or progesterone. MATERIAL AND METHODS: Sixty female rats were divided into six groups with 10 animals each: G1 (5 months) and GII (18 months) in the proestrus phase were treated with vehicle; the other animals were ovariectomized, and immediately after were treated with vehicle (GC) or with hormones ? (GE) treated with oestradiol benzoate (37.6 mg/animal); (GP) ? treated with medroxyprogesterone acetate (11.28 mg/animal) and (GEP) ? treated with oestradiol benzoate (37.6 mg/animal), more medroxyprogesterone acetate (11.28 mg/animal). In the GE group, the oestradiol was administered subcutaneously for seven days; in the GP group, the progestogen was administered subcutaneously for 23 dats; in the GEP group, the oestradiol was administered once a day in the first seven days and the progestogen throughout the next 23 days, subcutaneously. Twenty four hours after the last administration of hormones, the animals were euthanized, and the first pairs of inguinal breasts were removed, part of them being fixated in formaldehyde at 10% (morphological evaluation) and the other part submerged in acetone (evaluation of the glycosaminoglycans). The material destined to the morphology was included in paraffin, and the cuts were stained by hematoxylin-eosin. The material destined to the biochemistry was processed to the electrophoresis in agarose gel to sulfated GAGs analysis, and the hyaluronic acid was analyzed with a Elisa-Like fluorimetric method. The statistical analysis was made by the analysis of variance (ANOVA) followed by the Bonferroni test (p<0.05). RESULTS: The breasts of the groups with ovaries (GI and GII) as well as the ovariectomized control group (GC) have shown to be atrophic, wherein we observed typical alveoli with secretion in the lumen in the groups (GE) and (GEP); yet in the animals treated with progestogen (GP), we noticed alveoli formed basically by cells that occupied almost all the alveolar light. In relation to the data of the sulfated glycosaminoglycans we noticed higher concentration in the oophorectomized group (GC) due to high concentration of dermatan sulfate (GC = GE > GP = GEP > GII = GI; p<0.05). And in relation to the hyaluronic acid we found higher concentration in the animals of five months (GI > GII, GC, GE, GEP > GP; p<0.005).CONCLUSION: Oestradiol and progesterone had a trophic effect in the breast parenchyma. However, the administration of oestradiol beforehand changes the action of progesterone in the rat?s mammary tissue.
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    Alterações histoquímicas das glicosaminoglicanas na cérvice uterina no final da prenhez da rata albina após ministração local de hialuronidase
    (Federação Brasileira das Sociedades de Ginecologia e Obstetrícia, 2008-07-01) Lopes, Viviane Almeida de Alcântara [UNIFESP]; Luz, Mônica Regina Lourenço [UNIFESP]; Souza, Guilherme Negrão de [UNIFESP]; Fernandes Júnior, João Antão [UNIFESP]; Simões, Manuel de Jesus [UNIFESP]; Camano, Luiz [UNIFESP]; Souza, Eduardo de [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    PURPOSE: to study the histochemical changes related to the uterine cervix glycosaminoglycan of the albino female rat, after local ministration of hyaluronidases at the end of pregnancy. METHODS: ten female rats with positive pregnancy tests were randomly distributed in two numerically equal groups. The control group (Cg) was built up with rats that received a single dose of 1 mL of distilled water in the uterine cervix, under anesthesia, at the 18th pregnancy day. In the experimental group (Exg), the rats received 0.02 mL of hyaluronidase, diluted in 0.98 mL of distilled water (1 mL as a total), under the same conditions as the Cg. At the 20th pregnancy day, the rats were anesthetized once again and submitted to dissection, and the cervix prepared for histochemical study with alcian blue dye and its blockades (pH=0.5, pH=2.5, methylation and saponification). RESULTS: strongly positive reaction in the lamina propria (+3) has been seen in the Cg, and negative reaction in the Exg, with pH=0.5 alcian blue staining. With pH=2.5, staining has also been strongly positive (+4) in the Cg, and weakly positive (+1) in the Exg slide. After methylation, both groups have shown negative reaction, with pH=2.5 alcian blue staining. The lamina propria staining became negative after methylation in both groups, followed by saponification and enzymatic digestion on slide. CONCLUSIONS: there is clear predominance of sulphated glycosaminoglycans in the Cg as compared to the Exg and a small amount of identified carboxylated glycosaminoglycans in the Exg. The changes evidenced in the extracellular matrix have suggested that the hyaluronidase injected in the uterine cervix has promoted biochemical changes compatible with cervix maturation.
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    Análise histomorfológica e caracterização dos glicosaminoglicanos do útero de ratas tratadas com estrogênio, progestagênio e raloxifeno
    (Universidade Federal de São Paulo (UNIFESP), 2001) Simões, Manuel de Jesus [UNIFESP]
    Procurou-se avaliar, neste estudo, sob o ponto de vista histomorfomologico e bioquimico, os efeitos do tratamento com estrogenios conjugados equinos (CEE), isolados ou associados ao acetato de medroxiprogesterona (AMP), e do raloxifeno (RAL) sobre o endometrio de ratas adultas ooforectomizadas. Estudaram-se ratas adultas que, 60 dias apos serem castradas, foram distribuidas aleatoriamente, em cinco grupos: Grupo I - castrado; Grupo II tratadas com CEE (50 gg/kg, por dia); Grupo III - tratadas com AMP (0,2 mg/kg, por dia); Grupo IV - tratadas com CEE (50 gg/kg, por dia) + AMP (0,2 mg/kg, por dia); Grupo V - tratadas com RAL (3 mg/kg, por dia). A administracao dos farmacos foi feita por gavagem, sempre no periodo da manha, durante 28 dias consecutivos. Apos esse periodo os animais foram sacrificados e retiraram-se os uteros, os quais foram mergulhados em formol a 10 por cento. Em seguida, seccionaram-se os corpos uterinos, sendo a porcao media processada para estudo histologico e determinacao do indice de proliferacao nuclear (PCNA) dos epitelios superficial e glandular, em microscopia de luz; o restante do material foi processado para deteccao e determinacao bioquimica dos glicosaminoglicanos. Os resultados mostraram que os EEC isolados apresentaram efeito trofico sobre o endometrio e aumento significante do PCNA; a associacao de AMP inibiu, em parte, estes efeitos proliferativos. Ja nos grupos tratados com AMP ou RAL notamos atrofia endometrial e baixos indices de PCNA. Entretanto, no grupo que recebeu RAL identificamos a presenca de eosinofilos no estroma endometrial, o que sugere haver ligeira acao estrogenica. Os glicosaminoglicanos sulfatados (em especial o dermatam sulfato) apresentaram-se em niveis mais elevados nos animais tratados com EEC isolados ou associados ao AMP. Os indices de heparam sulfato mostraram-se baixos nos uteros dos animais tratados com AMP isoladamente ou associado aos EEC. A dosagem de acido hialuronico nao apresentou alteracoes entre os varios grupos de estudo. O raloxifeno nao alterou os niveis de glicosaminoglicanos no utero de ratas
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    Anti-inflammatory properties of a heparin-like glycosaminoglycan with reduced anti-coagulant activity isolated from a marine shrimp
    (Elsevier B.V., 2008-11-01) Brito, Adriana S.; Arimateia, Dayse S.; Souza, Lucilla R.; Lima, Marcelo A.; Santos, Vanessa O.; Medeiros, Valquiria P. [UNIFESP]; Ferreira, Paula A.; Silva, Rodrigo A.; Ferreira, Carmen V.; Justo, Giselle Z. [UNIFESP]; Leite, Edda L.; Andrade, Giulianna P. V.; Oliveira, Fernanda W.; Nader, Helena B. [UNIFESP]; Chavante, Suely F.; Univ Fed Rio Grande do Norte; Universidade Federal de São Paulo (UNIFESP); Universidade Estadual de Campinas (UNICAMP)
    The anti-inflammatory properties of a heparin-like compound from the shrimp Litopenaeus vannamei are related. Besides reducing significantly (p < 0.001) the influx of inflammatory cells to injury site in a model of acute inflammation, shrimp heparin-like compound was able to reduce the matrix metalloproteinase (MMPs) activity in the peritoneal lavage of inflamed animals. Moreover, this compound also reduced almost 90% the activity of MMP-9 secreted by human activated leukocytes. Negligible anti-coagulant activities in aPPT assay and a poor bleeding potential make this compound a better alternative than mammalian heparin as a possible anti-inflammatory drug. (C) 2008 Elsevier B.V. All rights reserved.
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    Avaliação audiológica em pacientes com mucopolissacaridose: estudo da ocorrência, tipo e grau de perda auditiva
    (Universidade Federal de São Paulo (UNIFESP), 2011-01-26) Silveira, Marcela Rosana Maia da [UNIFESP]; Azevedo, Marisa Frasson de [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Purpose: the mucopolysaccharidosis are a group of diseases caused by a deficiency in the lysosomal enzymes involved in the metabolism of mucopolysaccharides. Inherited metabolic diseases are caused by inborn errors of metabolism that lead to lack of proper functioning of certain enzymes. Hearing loss is a frequent manifestation in patients with mucopolysaccharidosis. The progressive nature of the hearing loss program requires audiologic, with periodic evaluations of the hearing. General objective: check the occurrence of hearing loss, characterizing its type and grade in patients with mucopolysaccharidosis. Specific objectives: aimed at verifying the possibility to evaluate patients with MPS and establish more effective procedures for hearing assessment in relation to age and type of MPS. Method: We evaluated 60 (sixty) patients, between 2 and 23 years, 25 females and 35 males with a diagnosis of different types of MPS, conducted by geneticist. Selected patients underwent audiometric assessment . The procedure for obtaining auditory thresholds were chosen according to the chronological age of the patient and also considering their position to understand / respond to the procedure. Results: Of the 60 patients studied, 53 (88.3%) completed the diagnostic audiology. Of these, 51 (96.2%) had hearing loss being the most conductive 25 (47.2%). There was no difference between the hearing thresholds by air and bone conduction thresholds logoaudiometric between the ears. There was a higher incidence of female patients in types I, III and IV of MPS and males in the types II and VI. Regarding the procedures used, pure tone audiometry was performed in patients over the types of MPS IV and VI. Type II patients performed more visual reinforcement audiometry. Sensorineural hearing loss occurred in patients with MPS type I, II and III. Type III patients had worse hearing thresholds obtained by bone and increased occurrence of type A tympanometric curves with the presence of acoustic reflex. Type IV patients showed a predominance of normal audiograms, with hearing thresholds better than others and increased occurrence of type A tympanometric curves. The type I showed more mixed hearing loss. There was a higher incidence of conductive hearing loss in type II, IV and VI of MPS with a higher incidence of tympanometric curves of type B and C in the absence of acoustic reflex. Conclusions: the audiological evaluation can be performed in 88,3% of the population; there was hearing loss in 96.2% of patients with MPS, with a predominantly conductive hearing loss of mild and moderately severe; was occurrence of sensorineural hearing loss in patients with MPS type I, II and III.
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    Avaliação do colágeno no osso de ratas diabéticas tipo 1 tratadas com isoflavonas
    (Universidade Federal de São Paulo, 2022-11-29) Vieira, Magno César [UNIFESP]; Simões, Manuel de Jesus [UNIFESP]; Carbone, Adriana Aparecida Ferraz [UNIFESP]; http://lattes.cnpq.br/9174521191304809; http://lattes.cnpq.br/5987164343458678; http://lattes.cnpq.br/5998705224419140
    Neste trabalho, o modelo “ratas diabéticas ovariectomizadas” foi utilizado pensando nas mulheres em menopausa e diabéticas com um tratamento alternativo com isoflavonas da soja, baseamo-nos em dados que mostram um aumento no número de mulheres que procuram utilizar-se de terapias hormonais. Objetivo: Analisar a matriz extracelular no osso de ratas diabéticas tratadas com isoflavonas ou 17β- estradiol. Métodos: Foram utilizadas 60 ratas (Rattus norvegicus albinus), fêmeas, adultas, ±3 meses de idade. Os animais foram separados em seis (6) grupos, a saber: controle Sham (n=10) animais não ovariectomizados na fase de estro; Sham+DM (n=10) controle Sham ratas diabéticas não ovariectomizadas na fase de estro; OVX (n=10) controle, ratas ovariectomizadas que receberam veículo propilenoglicol; OVX+DM (n=10) ratas diabéticas ovariectomizadas que receberam veículo propilenoglicol (fase de estro); OVX+DM+ISO (n=10) animais diabéticos ovariectomizados tratados com isoflavonas da soja (150mg/Kg, por gavagem); OVX+DM+E2 (n=10) animais diabéticos ovariectomizados tratados com estrogênio (17β-estradiol, 10μg/Kg, por via subcutânea). Para a indução do diabetes Tipo 1, as ratas receberam uma única injeção intraperitoneal de 60 mg/Kg de estreptozotocina (STZ, Sigma-Aldrich). O diabetes foi confirmado três dias após a injeção da estreptozotocina. Todos os animais foram tratados durante 30 dias consecutivos, e ao final, os animais foram anestesiados e os fêmures removidos e processados para estudo em H.E. e Picro Sirius Red. Para a análise dos resultados foi utilizado Oneway ANOVA seguido do teste de Tukey. Foram considerados estatisticamente significativos experimentos cujo valor de p foi menor que 5% (p ≤ 0,05) para significância estatística. Os cálculos foram feitos com o programa SPSS versão13 (SPSS, Chicago, IL). Resultados: Os resultados obtidos foram os seguintes: Peso corporal: não foi diferente entre os grupos não diabéticos Sham e OVX e, os grupos de ratas diabéticas Sham+DM, OVX+DM, OVX+DM+ISO e OVX+DM+E2 (p>0,05). No entanto, houve uma diminuição significativa do peso corporal nos grupos de ratas diabéticas em comparação com os grupos não diabéticas Sham e OVX (p<0,001). Sensibilidade à Insulina: Os valores mais baixos de insulina tolerância foram observadas no OVX+DM (2,41±0,95). Os grupos Sham (4,64±0,95) e OVX (4,57±0,58) apresentaram os maiores valores de sensibilidade à insulina (p<0,001). x Os grupos OVX+DM+ISO e OVX+DM+E2 apresentaram valores semelhantes, inferiores aos Sham e OVX e superior ao grupo OVX+DM. Histomorfometria e Análise Bioquímica de Glicosaminoglicanos Sulfatados: O volume do osso trabecular foi maior nos grupos de ratas diabéticas ovariectomizadas OVX+DM+E2 (26,35±2,13) e OVX+DM+ISO (18,36±173), tratadas, respectivamente, com 17β estradiol e isoflavonas e menor no OVX+DM (8,32±2,52), grupo de ratas diabéticas ovariectomizadas que receberam veículo propilenoglicol em fase de estro. Resultados semelhantes foram encontrados em relação à espessura do osso cortical, que foi maior no OVX+DM+E2 (398,4±1,74) e OVX+DM+ISO (295,6±1,45) e menor no OVX+DM (142,6±2,74) (p<0,05). Com relação ao sulfato de condroitina foi encontrado nos ossos de todos os grupos de animais estudados. Picro Sirius Red: As fibras colágenas de substância óssea trabecular e cortical das epífises dos fêmures apresentaram maior intensidade de birrefringência nos grupos (OVX+DM+ISO 330,2±2 e OVX+DM+E2 414,8± 33,7) tratados, respectivamente, com isoflavonas e 17β estradiol
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    Avaliação prostpectiva de heparam sulfato, condroitim sulfato e ácido hialurônico em pacientes com câncer de próstata
    (Universidade Federal de São Paulo (UNIFESP), 2018-06-28) Silva, Matheus Neves Ribeiro Da [UNIFESP]; Pinhal, Maria Aparecida Da Silva [UNIFESP]; http://lattes.cnpq.br/7511274763693292; http://lattes.cnpq.br/8143551334488274; Universidade Federal de São Paulo (UNIFESP)
    Prostate cancer in Brazil represents the most common type of cancer among men after skin cancer. Although there are effective tests for the diagnosis of prostate cancer, there are still discussions about the most appropriate and non-invasive method for such diagnosis, always looking for less invasive methods to evaluate the possible prognosis of the disease. The present study aimed to evaluate urinary levels of chondroitin sulfate (CS) and heparan sulfate (HS), as well as plasma levels of hyaluronic acid (HA) in patients with prostate cancer, before and after treatment, compared with individuals which was not affected by neoplasia. Plasma samples were used to quantify AH and urine for CS and HS quantification from forty-four patients with prostate cancer and fourteen controls (non-affected individuals). Laboratory and pathological clinical data were correlated with the quantification of glycosaminoglycans by statistical analysis. There was no difference in the urinary levels of CS and HS among patients with prostate cancer and the control group. However, sulfated glycosaminoglycans (HS and CS) were analyzed in urine samples collected before and after treatment, and an increased in urinary excretion of heparan sulfate was observed after surgery, followed by decrease in the excretion of chondroitin sulfate after hormone therapy. The level of serum hyaluronic acid showed a significant increase in patients with prostate cancer (39.68 ± 30.00 ng / ml), compared to the control group (15.04 ± 7.11 ng / ml), p = 0.004. We also observed a significant increase in hyaluronic acid in patients with high-risk of prostate cancer, following the D'Amico classification, compared to low-risk patients (p = 0.0214). Patients submitted to surgery had a significant reduction in the level of AH (p = 0.029). The quantification of serum hyaluronic acid represents a noninvasive test with potential use in the diagnostic and prognostic evaluation of prostate cancer. Future studies are essential to confirm the unprecedented but preliminary results obtained in the present study.
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    Caracterização de glicosaminoglicanos urinários de pacientes portadores de mucopolissacaridose do tipo IV A
    (Universidade Federal de São Paulo (UNIFESP), 2006) Coulson-Thomas, Vivien Jane [UNIFESP]; Toma, Leny [UNIFESP]
    A Mucopolissacaridose do tipo IVA (MPS IVA) é uma doença genética do tecido conjuntivo, que envolve mutações no gene N-acetil galactosamina-6-sulfato sulfatase. Esta enzima participa da degradação seqüencial do condroitim sulfato (CS) e queratam sulfato (KS), removendo sulfato da N-acetilgalactosamina-6-sulfato e galactose–6-sulfato do terminal não redutor destes GAGs. Quando esta enzima está ausente, os órgãos acumulam produtos de digestão incompleta destes GAGs, levando a um aumento na excreção de tais compostos na urina. Neste estudo nós analisamos KS e CS purificados de urina doada por pacientes portadores de MPS IVA e de indivíduos normais. GAGs totais foram extraídos da urina de indivíduos normais e pacientes utilizando uma resina de troca iônica . O KS foi isolado, incubando GAGs totais com extrato bruto de enzimas da F. heparinum, que digere todos os GAGs, com exceção do KS. Os produtos menores remanescentes foram separados do composto intacto por cromatografia de gel filtração sequenciais, Sephadex G-25 (PD10) e Sephadex G-10. KS foi detectado através de eletroforese em poliacrilamida e confirmado com queratanases específicas. Na tentativa de separar KS e CS, mantendo a integridade de ambos, os GAGs totais foram submetidos a cromatografia em Sephadex G-50 para fracionamento de acordo com a massa molecular. O perfil de eluição de CS foi detectado com reagente de carbazol que reage com ácido urônico, presente no CS e não no KS. Este foi detectado nas frações através de blotting com o anticorpo monoclonal específico para queratam sulfato. A maior parte do KS foi detectado na membrana de nitrocelulose, indicando um possível KS-ligado a peptídeo na urina. O mesmo procedimento de blotting foi realizado, e a membrana de nitrocelulose corado com reagente Ponceau, revelando a presença de proteína comigrando com o KS previamente detectado. Para confirmar o complexo KS-peptídeo GAGs totais foram submetidos a cromatografia hidrofóbica em coluna de Octyl-Sepharose, e eluídos com um gradiente linear de sulfato de amônio 2M e uma lavagem final com isopropanol50%. Uma população de KS foi retida na coluna, sendo eluído apenas com isopropanol 50%, devido à interação hidrofóbica do peptídeo com a resina. KS, CS e proteína foram detectados nessa população retida. Amostras de GAGs totais foram submetidos a uma reação química de β-eliminação, que remove açucares O-ligados a proteínas, e então aplicados a HPLC, em colunas de fase reversa, C4 Ultrasphere rotein and C18 Sephasil. Vários fragmentos protéicos ou peptídicos foram purificados nas colunas, que estão sendo sequenciados para análise estrutural..
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    Connexin37 deficiency alters organic bone matrix, cortical bone geometry, and increases Wnt/beta-catenin signaling
    (Elsevier Science Inc, 2017) Pacheco-Costa, Rafael [UNIFESP]; Kadakia, Jay R.; Atkinson, Emily G.; Wallace, Joseph M.; Plotkin, Lilian I.; Reginato, Rejane D. [UNIFESP]
    Deletion of connexin (Cx) 37 in mice leads to increased cancellous bone mass due to defective osteoclast differentiation. Paradoxically
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    A dose-optimization trial of laronidase (Aldurazyme (R)) in patients with mucopolysaccharidosis I
    (Elsevier B.V., 2009-01-01) Giugliani, Roberto; Rojas, Veronica Munoz; Martins, Ana Maria [UNIFESP]; Valadares, Eugnia R.; Clarke, Joe T. R.; Goes, Jose E. C.; Kakkis, Emil D.; Worden, Mary Alice; Sidman, Marisa; Cox, Gerald F.; Univ Fed Rio Grande do Sul; Universidade Federal de São Paulo (UNIFESP); Universidade Federal de Minas Gerais (UFMG); Hosp Sick Children; Hosp Infantil Joana de Gusmao; BioMarin Pharmaceut Inc; Genzyme Corp; Harvard Univ
    Recombinant human alpha-L-iduronidase (Aldurazyme (R) laronidase) is approved as an enzyme replacement therapy to treat the lysosomal storage disorder, mucopolysaccharidosis type I (MPS I) at a dose of 0.58 mg/kg by once-weekly intravenous infusion. To assess whether alternate dosing regimens might provide a better reduction in lysosomal storage, a 26-week, randomized, open-label, multinational dose-optimization trial was conducted. the pharmacodynamic effect and safety of the approved laronidase dose was compared to three alternative regimens (1.2 mg/kg every 2 weeks; 1.2 mg/kg every week; 1.8 mg/kg every 2 weeks) among 33 MPS I patients. the four treatment regimens showed no significant differences in the reduction of urinary glycosaminoglycan excretion or liver volume. Laronidase had an acceptable safety profile in all dose regimen groups. Infusion-associated reactions were the most common drug-related adverse events across dose regimens (by patient incidence), and included pyrexia (21%), vomiting (15%), rash (15%), and urticaria (12%). Patients in the approved dose group had the lowest incidence of drug-related adverse events (38% vs. 63-75%) and infusion-associated reactions (25% vs. 25-63%). There was one death: a patient with acute bronchitis died of respiratory failure 6h after completing the first laronidase infusion. the approved 0.58 mg/kg/week laronidase dose regimen provided near-maximal reductions in glycosaminoglycan storage and the best benefit-to-risk ratio. the 1.2 mg/kg every 2 weeks regimen may be an acceptable alternative for patients with difficulty receiving weekly infusions, but the long-term effects of this regimen are unknown. (C) 2008 Elsevier Inc. All rights reserved.
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    Efeito dos glicosaminoglicanos na adesão e internalização celular de nanopartículas de poliestireno
    (Universidade Federal de São Paulo (UNIFESP), 2021) Olivieri Junior, Paulo Henrique [UNIFESP]; Sousa, Alioscka Augusto Caldeira Araujo [UNIFESP]; Universidade Federal de São Paulo
    Understanding the factors that regulate cellular endocytosis of synthetic nanoparticles is an important objective in many fields of science and technology, for example, in nanomedicine in applications covering drug delivery. However, despite decades of study in the field, predicting (or even explaining) specific results remains a challenge. This is not surprising, considering the many factors that can influence nanoparticle– cell interactions. One of these factors concerns the negatively charged glycosaminoglycans (GAGs) that integrate the cellular glycocalyx. Despite its importance, so far only a few studies have examined the impact of GAGs on cellular internalization of nanoparticles. Therefore, the main objective of this work was to understand the influence of surface GAGs in the internalization process of nanoparticles. Anionic and cationic polystyrene nanoparticles of 50 nm in diameter were used as a model system. Wild-type CHO (CHOK1) and mutant (CHO-745, deficient in GAG synthesis) cells were used as a model cell line. The influence of the specific GAGs heparan and chondroitin sulfates (HS and CS) on particle internalization was also evaluated through the selective enzymatic removal of these GAGs in the wild-type cells. All experimental steps were meticulously standardized to minimize variability between experiments. The results showed that the surface GAGs acted as an electrostatic filter, hindering the uptake of negatively charged particles, while enhancing the uptake of those with positive charge. Data obtained from the selective removal of HS and CS indicated that the 'electrostatic filter' function was performed mainly by HS, which is in accordance with its greater negative charge density relative to CS.
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    Estudo comparativo da expressão de glicosaminoglicanos, fibronectina e adesão celular em culturas de fibroblastos gengivais normais e de pacientes com fibromatose gengival hereditária. Efeito da Ciclosporina A
    (Universidade Federal de São Paulo (UNIFESP), 2003) Dreyfuss, Juliana Luporini [UNIFESP]; Nader, Helena Bonciani [UNIFESP]
    Fibromatose gengival hereditaria (FGH) e fibromatose gengival induzida por ciclosporina sao hiperplasias caracterizadas por um aumento volumetrico da' gengiva. Doenca de origem fibrotica progressiva, benigna e nao-hemorragica, que pode envolver tanto a maxila quanto a mandibula (Tipton, 1997). Alguns estudos demonstram a sintese aumentada de componentes da matriz extracelular tais como fibronectina e colagenos (Shirasuna et al.,1988). Os glicosaminoglicanos (GAGs) constituem um dos principais componentes do tecido conjuntivo gengival. No', presente estudo, foi analisada a expressao dos glicosaminoglicanos por fibroblastos em cultura de pacientes portadores de FGH (membros da mesma familia), bem como fibroblastos de gengiva normal (GIM). Como o tratamento com a ciclosporina,' droga imunossupressora, induz fibromatose gengival, fibroblastos gengivais normais foram tratados em cultura com 200ng/mL de ciclosporina A (GNc) para avaliacao do padrao de expressao dos GAGs. Os glicosaminoglicanos sulfatados foram analisados na 4a e na 8' passagem, nos diferentes compartimentos celulares: meio de cultura, superficie celular e fracao celular (celula+matriz extracelular) atraves da marcacao metabolica com (35S]-sulfato de sodio (l50gCilmL) e posterior analise por eletroforese em gel de agarose. Os tipos de dissacarideos formados pela acao das enzimas condroitinases AC e ABC foram tambem determinados. O acido hialuronico' (AH) foi dosado atraves do metodo fluorimetrico ELISA-like. O dermatam sulfato' (DS), condroitim sulfato (CS), heparam sulfato (HS) e AH estao presentes em todos os compartimentos celulares em todos os fibroblastos estudados. Nao obstante, as, quantidades relativas e absolutas variam de acordo com o compartimento. Alem disso, os fibroblastos FGH e GNc demonstraram um padrao de distribuicao dos GAGs sulfatados diferente quando comparados a GN. Ocorre uma expressiva diminuicao da sintese de GAGs sulfatados, e um aumento de ate 6 vezes na sintesea(au)
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    Estudo dos glicosaminoglicanos aórticos: sua importância funcional e interação com lipoproteinas plasmáticas
    (Universidade Federal de São Paulo (UNIFESP), 1981) Toledo, Olga Maria Szymanski [UNIFESP]; Mourão, Paulo Antonio de Souza [UNIFESP]
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    Evaluation of the metabolism of glycosaminoglycans in patients with interstitial cystis
    (Sociedade Brasileira de Urologia, 2014-01-01) Lucon, Marcos; Martins, Joao Roberto; Leite, Kátia Ramos Moreira [UNIFESP]; Soler, Roberto; Nader, Helena Bonciani [UNIFESP]; Srougi, Miguel [UNIFESP]; Bruschini, Homero; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    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.
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    Expressão de glicosaminoglicanos e proteínas da matriz extracelular em células endoteliais
    (Universidade Federal de São Paulo (UNIFESP), 2008) Dominato, Juliana Augusta Albieri [UNIFESP]; Nader, Helena Bonciani [UNIFESP]
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    Expressão de glicosaminoglicanos sulfatados e proteoglicanos de heparam sulfato em células endoteliais resistentes ao anoikis após tratamento com trastuzumabe
    (Universidade Federal de São Paulo (UNIFESP), 2017) Onyeisi, Jessica Oyie Sousa [UNIFESP] ; Azevedo, Carla Cristina Lopes de [UNIFESP]; http://lattes.cnpq.br/6280504619546509; http://lattes.cnpq.br/0546695375822929; Universidade Federal de São Paulo (UNIFESP)
    Objetivo: Avaliar o efeito do trastuzumabe (Tmab) na expressão de glicosaminoglicanos sulfatados e proteoglicanos de heparam sulfato em células endoteliais resistentes ao anoikis. Métodos: Células endoteliais de aorta de coelho (EC), EC resistentes ao anoikis (Adh-EC) e EC transfectada com o oncogene EJ-ras (EJ-ras EC) foram tratadas com trastuzumabe e analisadas em relação a: capacidade adesiva, invasão, proliferação, angiogênese, ciclo celular, expressão e localização do sindecam-4, expressão de perlecam e síntese de glicosaminoglicanos sulfatados (GAGs). Resultados: Os resultados demonstraram que as células endoteliais anoikis-resistentes apresentam maior adesão à fibronectina e menor capacidade invasiva, proliferativa e angiogênica, após tratamento com trastuzumabe. Observamos também que, após o tratamento com o trastuzumabe, houve um aumento significativo no número de células na fase S do ciclo celular. Em relação à expressão do sindecam-4, perlecam e síntese de heparam sulfato (HS), observamos uma diminuição tanto na expressão do sindecam-4 e perlecam, bem como na biossíntese de HS nas células endoteliais resistentes ao anoikis após exposição ao Tmab. Por outro lado, as células endoteliais selvagens apresentaram aumento significativo na expressão de perlecam e na síntese de HS. Conclusão: Juntos, os resultados sugerem que o trastuzumabe não interage apenas com HER2, mas com glicosaminoglicanos e proteoglicanos tanto de superfície celular, como da matriz extracelular, e também, controla eventos celulares das células endoteliais resistentes ao anoikis.
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    Expressão dos glicosaminoglicanos e proteoglicanos na matriz extracelular da cérvice uterina da rata albina prenhe após injeção local de hialuronidase
    (Universidade Federal de São Paulo (UNIFESP), 2012) Souza, Guilherme Negrão de [UNIFESP]; Camano, Luiz [UNIFESP]
    Objetivo: Avaliar a expressao dos glicosaminoglicanos (GAGs) e proteoglicanos (PGs) na matriz extracelular (MEC) da cervice uterina da rata albina prenhe, apos injecao local de hialuronidase. Metodologia: Dez ratas prenhes foram distribuidas aleatoriamente em dois grupos, numericamente iguais. O grupo controle (Gc) foi constituido pelas ratas que receberam 1 mL de agua destilada, dose unica, no 18º dia da prenhez, sob anestesia, ministrado na cervice uterina. No grupo experimental (Gexp), os animais receberam, sob as mesmas condicoes do Gc, 0,02 mL de hialuronidase, diluido em 0,98 ml de agua destilada (total de 1ml). No 20º dia de prenhez, seis horas apos marcacao in vivo de [35SO4] de sulfato radioativo, as ratas foram sacrificadas por inalacao de monoxido de carbono e submetidas a disseccao, preparando-se a cervice uterina para caracterizacao e quantificacao dos GAGs recemsintetizados, condroitim sulfato (CS), dermatam sulfato (DS) e heparam sulfato (HS), extraidos apos proteolise e determinados e caracterizados, apos eletroforese em gel de agarose, por densitometria optica, antes e apos degradacao enzimatica com enzimas especificas. Foram avaliadas tambem a expressao qualitativa do acido hialuronico (AH) por histoquimica e sua quantificacao por ensaio tipo sanduiche em placa de ELISA. A analise estatistica de todas as informacoes coletadas nesta pesquisa foi inicialmente feita de forma descritiva. Para as variaveis de natureza quantitativa (numerica) foram calculadas mediana, valor minimo, valor maximo e confeccionados graficos do tipo diagrama de dispersao unidimensional. A analise inferencial, empregada com o intuito de comparar os valores da quantidade dos GAGs entre os grupos, foi o Teste t-Student para amostras independentes, com o nivel de significancia α igual a 5%. Tambem, nesse estudo, diferentes PGs foram avaliados por imunofluorescencia com anticorpos. Resultados: O comportamento da sintese destas moleculas ocorreu de forma similar e a degradacao enzimatica confirmou a presenca de CS/DS e HS na composicao das amostras estudadas; houve diminuicao significante na quantidade total dos GAGs sulfatados e do AH no Gexp se comparado ao Gc. Na histoquimica, ha dispersao na distribuicao do AH no Gexp. Paralelamente, observou-se no Gexp menor expressao (imunofluorescencia) de diversos PGs como o decorim, biglicam, sindecam e CD44 Conclusoes: O tratamento com hialuronidase (Gexp) promove queda significativa da concentracao dos GAGs sulfatados na cervice uterina de ratas albinas prenhes. Tanto a expressao qualitativa (por histoquimica) quanto a quantitativa (dosagem fluorometrica) do AH tambem se mostraram reduzidas no Gexp. O conjunto dos resultados permite inferir que as alteracoes desses compostos na cervice uterina no Gexp, comparado ao Gc, promovem modificacoes locais semelhantes ao processo de maturacao cervical fisiologica, fatores primordiais ao desencadeamento da parturicao
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    Expression of heparanase in basal cell carcinoma and squamous cell carcinoma
    (Soc Brasileira Dermatologia, 2016) Silva Pinhal, Maria Aparecida [UNIFESP]; Leal Almeida, Maria Carolina [UNIFESP]; Costa, Alessandra Scorse [UNIFESP]; Theodoro, Therese Rachell [UNIFESP]; Serrano, Rodrigo Lorenzetti [UNIFESP]; Santos Machado Filho, Carlos D'Apparecida [UNIFESP]
    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.
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    Extracellular matrix alterations in the Peyronie's disease
    (Elsevier Science Bv, 2017) Watanabe, Marcelo Silva; Theodoro, Therese Rachel; Coelho, Natalia Lima [UNIFESP]; Mendes, Aline [UNIFESP]; Pereira Leonel, Monica Luzia; Mader, Ana Maria; Nader, Helena Bonciani [UNIFESP]; Glina, Sidney; Silva Pinhal, Maria Aparecida [UNIFESP]
    Peyronie's disease is characterized by fibrous plaque formation of the tunica albuginea, causing penile deformity and fertility problems. The aim of the present study was to investigate alterations in the extracellular matrix in Peyronie's disease. The study used tissues collected by surgical procedure from individuals that presented a well-established disease, while control samples were obtained by biopsies of fresh cadavers. Immunohistochemistry analysis followed by digital quantification was performed to evaluate TGF-beta, heparanases and metalloproteinases (MMPs). The profile of sulfated glycosaminoglycans, chondroitin sulfate and dermatan sulfate was determined by agarose gel electrophoresis, while hyaluronic acid quantification was obtained by an ELISA-like assay. The expression of mRNA was investigated for syndecan-1 proteoglycan (Syn-1), interleukine-6 (IL-6), hyaluronic acid synthases, and hyaluronidases. Pathologic features showed decreased apoptosis and blood vessel number in Peyronie's tissues. TGF-beta and IL-6 were significantly enhanced in Peyronie's disease. There was an increased expression of heparanases, though no alteration was observed for MMPs. Hyaluronic acid as well as hyaluronic acid synthases, hyaluronidases, and dermatan sulfate were not changed, while the level of chondroitin sulfate was significantly (P = 0.008, Mann-Whitney test) increased in Peyronie's samples. Heparanases and sulfated glycosaminoglycans seem to be involved in extracellular matrix alterations in Peyronie's disease. (C) 2017 Production and hosting by Elsevier B.V. on behalf of Cairo University.