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1.
Arq. bras. oftalmol ; 83(5): 378-382, Sept.-Oct. 2020. graf
Article in English | LILACS | ID: biblio-1131627

ABSTRACT

ABSTRACT Purpose: To measure humor heat-shock protein 70, periostin, and irisin levels in patients with pseudoexfoliation syndrome and cataract (without glaucoma), and compare them with those of patients with cataract but without pseudoexfoliation. Methods: We examined 31 eyes of 31 patients with pseudoexfoliation and cataract (without glaucoma) and 30 eyes of 30 patients with cataract. We collected aqueous humor samples from all patients at the time of cataract surgery through a limbal paracentesis via a 25-gauge cannula mounted on a tuberculin syringe that received 100 to 150 µL of aqueous humor. We measured levels of aqueous humor Heat shock protein 70, periostin, and irisin using enzyme-linked immunosorbent assay methods. Results: The age (p=0.221) and gender (p=0.530) means were similar between the pseudoexfoliation and control groups. The mean Heat shock protein 70 level (29.22 ± 9.46 ng/mL; 17.88-74.46) in the pseudoexfoliation group was significantly higher than that in the control group (19.03 ± 7.05 ng/mL; 9.93-35.52; p<0.0001). The mean periostin level was significantly higher (6017.32 ± 1271.79 pg/mL; 3787.50-10803.57) in the pseu doexfoliation group than that in the control group (4073.63 ± 1422.79 pg/mL; 2110.44-7490.64; p<0.0001). The mean irisin level (53.77 ± 10.19 ng/mL; 29.46-71.16) was significantly higher than that in the control group (39.29 ± 13.58 ng/mL; 19.41-70.56; p<0.0001). Conclusions: Heat shock protein 70, periostin, and irisin levels increase in the aqueous humor of patients with pseudoexfoliation without glaucoma.


RESUMO Objetivo: Comparar os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso de pacientes com pseudoexfoliação com catarata sem glaucoma e compará-los com pacientes com catarata sem pseudoexfoliação. Métodos: Trinta e um olhos de 31 pacientes com pseudoexfoliação com catarata sem glaucoma e 30 olhos de 30 indivíduos com catarata foram incluídos neste estudo. Amostras de humor aquoso foram coletadas de todos os pacientes no momento da cirurgia de catarata e obtidas através de uma paracentese límbica por meio de uma cânula de calibre 25 acoplada a uma seringa com tuberculina. Foram coletados 100 a 150 µL de humor aquoso. Os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso foram medidos usando o método de ensaio imunossorvente ligado a enzima. Resultados: A média da idade (p=0,221) e sexo (p=0,530) foram semelhantes entre os grupos pseudoexfoliação e controle. Os níveis médios de proteína de choque térmico 70 foram 29,22 ± 9,46 ng/mL (17,88-74,46) e 19,03 ± 7,05 ng/ mL (9,93-35,52) nos grupos pseudoexfoliação e controle, respectivamente. Os níveis de proteína de choque térmico 70 foram maiores no grupo pseudoexfoliação (p<0,0001). O nível médio de periostina foi de 6017,32 ± 1271,79 pg/mL (3787,50-10803,57) no grupo pseudoexfoliação e 4073,63 ± 1422,79 pg/mL (2110,44-7490,64) no grupo controle. O nível médio de periostina também foi maior no grupo pseudoexfoliação (p<0,0001). Os níveis médios de irisina foram 53,77 ± 10,19 ng/mL (29,46-71,16) e 39,29 ± 13,58 ng/mL (19,41-70,56) nos grupos pseudoexfoliação e controle, respectivamente. O nível médio de irisina foi maior no grupo pseudoexfoliação do que no grupo controle (p<0,0001). Conclusões: Os níveis de proteína de choque térmico 70, de periostina e de irisina aumentam no humor aquoso de pacientes com pseudoexfoliação sem glaucoma.


Subject(s)
Humans , Aqueous Humor , Cataract , Cell Adhesion Molecules , Glaucoma , Fibronectins , Exfoliation Syndrome , HSP70 Heat-Shock Proteins , Enzyme-Linked Immunosorbent Assay , Cell Adhesion Molecules/metabolism , Fibronectins/metabolism , Exfoliation Syndrome/metabolism , HSP70 Heat-Shock Proteins/metabolism
2.
Acta cir. bras ; 34(1): e20190010000007, 2019. tab, graf
Article in English | LILACS | ID: biblio-983684

ABSTRACT

Abstract Purpose: To investigate the impact of Ramipril (RAM) on the expressions of insulin-like growth factor-1 (IGF-1) and renal mesangial matrix (RMM) in rats with diabetic nephropathy (DN). Methods: The Sprague Dawley rats were divided into normal control (NC) group (n = 12), DN group (n = 11), and DN+RAM group (n = 12). The ratio of renal weight to body weight (RBT), fasting blood glucose (FBG), HbA1c, 24-h urine protein (TPU), blood urea nitrogen (BUN), creatinine (Cr), renal pathological changes, the levels of IGF-1, fibronectin (FN), type IV collagen (Col-IV), and matrix metalloproteinases (MMP)-2 were compared among the groups. Results: Compared with NC group, the RBT, FBG, HbA1c, TPU, BUN, Cr, and RMM in DN group were significantly increased (P < 0.05), the IGF-1, FN, and Col-IV were significantly upregulated (P < 0.05), while MMP was significantly downregulated (P < 0.05). Compared with DN group, the indexes except for the FBG and HbA1c in DN+RAM group were significantly improved (P < 0.05), among which IGF-1 exhibited significant positive correlation with TPU(r=0.937), FN(r=0.896) and Col-IV(r=0.871), while significant negative correlation with MMP-2 (r=-0.826) (P<0.05). Conclusion: RAM may protect the kidneys by suppressing IGF-1 and mitigating the accumulation of RMM.


Subject(s)
Animals , Male , Rats , Insulin-Like Growth Factor I/antagonists & inhibitors , Angiotensin-Converting Enzyme Inhibitors/pharmacology , Ramipril/pharmacology , Diabetic Nephropathies/drug therapy , Mesangial Cells/drug effects , Insulin-Like Growth Factor I/metabolism , Immunohistochemistry , Fibronectins/drug effects , Fibronectins/metabolism , Rats, Sprague-Dawley , Matrix Metalloproteinases/drug effects , Matrix Metalloproteinases/metabolism , Collagen Type IV/adverse effects , Collagen Type IV/metabolism , Diabetic Nephropathies/metabolism , Mesangial Cells/metabolism
4.
Arch. endocrinol. metab. (Online) ; 61(4): 382-390, July-Aug. 2017. tab, graf
Article in English | LILACS | ID: biblio-887578

ABSTRACT

ABSTRACT Objective Patients infected with the human immunodeficiency virus (HIV) have an increased risk of metabolic disorders and alterations on irisin levels. Therefore, the purpose of the current investigation was to quantify the circulating irisin concentration in HIV-infected subjects under highly active antiretroviral therapy and to determine possible correlations between irisin levels with fat mass, fat-free mass, body mass index (BMI), and muscle strength. Subjects and methods Cross-sectional study of 10 men (36.7 ± 11.3 years) and 10 women (42.5 ± 10.3 years) infected with HIV, recruited from the Specialized Service Center in the State Center of Reference for High and Medium Complexity. Blood samples were collected to determine plasma irisin levels, glucose, HDL, total cholesterol, triglycerides, and LDL. Body composition (fat mass, fat-free mass) and anthropometrics (body mass index; BMI) were measured by bioelectrical impedance. Muscle strength was assessed using a mechanic hand dynamometer and one maximum repetition tests. Results Irisin levels correlated positively with fat mass (r = 0.67; p = 0.001) and BMI (r = 0.48; p = 0.036). In contrast, there was an inverse correlation between irisin levels and fat-free mass (r = -0.41; p = 0.008) and five strength parameters: right hand grip (r = -0.46; p = 0.044); left hand grip (r = -0.50; p = 0.027), relative hand grip (r = -0.79; p = 0.001), bench press (r = -0.58; p = 0.009), leg press (r = -0.40; p = 0.085), and biceps curl (r = -0.059; p = 0.009). Conclusion Irisin levels correlated positively with body fat and negatively with fat-free mass and strength parameters in HIV-infected patients. Female patients infected with HIV receiving highly active antiretroviral therapy have higher levels of irisin compared with men in a similar circumstance.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , HIV Infections/blood , Adipose Tissue/drug effects , Adipose Tissue/pathology , Fibronectins/blood , Body Composition/drug effects , HIV Infections/metabolism , HIV Infections/drug therapy , Sex Factors , Cross-Sectional Studies , Fibronectins/metabolism , Fibronectins/pharmacology , Hand Strength , Antiretroviral Therapy, Highly Active , Anti-Retroviral Agents/therapeutic use , Muscle Strength/drug effects
5.
Acta cir. bras ; 32(5): 350-358, May 2017. tab, graf
Article in English | LILACS | ID: biblio-837705

ABSTRACT

Abstract Purpose: To investigate the mechanisms by which PD98059 and LY294002 interfere with the abnormal deposition of extracellular matrix regulated by connective tissue growth factor (CTGF) of rat pulmonary artery smooth muscle cells (PASMCs). Methods: Rat PASMCs were cultured and separated into a control group. Real-time fluorescence quantitative PCR was performed to detect the expression of collagen III and fibronectin mRNA. Immunohistochemistry and western blot analyses were performed to detect the expression of collagen III protein. Results: The expression of collagen III and fibronectin mRNA was greater in PASMCs stimulated with CTGF for 48 h, than in the control group. After 72h of stimulation, the expression of collagen III protein in the PASMCs was greater than in the control. The equivalent gene and protein expression of the CPL group were much more significant. Conclusions: CTGF can stimulate the gene expression of collagen III and fibronectin in PASMCs, which may be one of the factors that promote pulmonary vascular remodeling (PVR) under the conditions of pulmonary arterial hypertension (PAH). PD98059 and LY294002 can inhibit the ERK1/2 and PI3K/PKB signaling pathways, respectively, thus interfering with the biological effects of CTGF. This may be a new way to reduce PAH-PVR.


Subject(s)
Animals , Male , Flavonoids/pharmacology , Chromones/pharmacology , Fibronectins/metabolism , MAP Kinase Signaling System/drug effects , Collagen Type III/metabolism , Connective Tissue Growth Factor/pharmacology , Pulmonary Artery/cytology , Gene Expression/drug effects , Cells, Cultured , Gene Expression Regulation , Fibronectins/genetics , Rats, Sprague-Dawley , Phosphatidylinositol 3-Kinases/metabolism , Models, Animal , Collagen Type III/genetics , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Connective Tissue Growth Factor/metabolism
6.
Clinics ; 71(6): 325-331, tab, graf
Article in English | LILACS | ID: lil-787427

ABSTRACT

OBJECTIVES: We evaluated mRNA expression levels of genes that encode TGF-β1; the TGF-β1 receptor; the collagen-modifying enzymes LOX, PLOD1, and PLOD2; and the extracellular matrix proteins COMP, FN1, TNC and TNXB in synovial/capsule specimens from patients with idiopathic adhesive capsulitis. Possible associations between the measured mRNA levels and clinical parameters were also investigated. METHODS: We obtained glenohumeral joint synovium/capsule specimens from 9 patients with idiopathic adhesive capsulitis who had not shown improvement in symptoms after 5 months of physiotherapy. Adhesive capsulitis was confirmed in all patients by magnetic resonance imaging. We also obtained specimens from 8 control patients who had underwent surgery for acute acromioclavicular joint dislocation and who had radiological indication of glenohumeral capsule alteration based on arthroscopic evaluation. mRNA expression in the synovium/capsule specimens was analyzed by quantitative reverse transcription PCR. The B2M and HPRT1 genes were used as references to normalize target gene expression in the shoulder tissue samples. RESULTS: The synovium/capsule samples from the patients with adhesive capsulitis had significantly higher TNC and FN1 expression than those from the controls. Additionally, symptom duration directly correlated with expression of TGFβ1 receptor I. CONCLUSION: Elevated levels of TNC and FN1 expression may be a marker of capsule injury. Upregulation of TGFβ1 receptor I seems to be dependent on symptom duration; therefore, TGFβ signaling may be involved in adhesive capsulitis. As such, TNC, FN1 and TGFβ1 receptor I may also play roles in adhesive capsulitis by contributing to capsule inflammation and fibrosis.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Bursitis/metabolism , Fibronectins/metabolism , Shoulder Joint/metabolism , Synovial Membrane/metabolism , Tenascin/metabolism , Transforming Growth Factor beta1/genetics , Acromioclavicular Joint/injuries , Acromioclavicular Joint/metabolism , Bursitis/genetics , Case-Control Studies , Extracellular Matrix Proteins/metabolism , Gene Expression , Joint Dislocations/metabolism , Pilot Projects , RNA, Messenger/metabolism , Transforming Growth Factor beta1/metabolism
7.
Rev. bras. epidemiol ; 18(1): 248-261, Jan-Mar/2015. tab, graf
Article in Portuguese | LILACS | ID: lil-736433

ABSTRACT

OBJETIVO: Analisar as desigualdades socioeconômicas na utilização de consultas médicas (CM) no último ano no Brasil. MÉTODOS: Dados da Pesquisa Nacional por Amostra de Domicílios (≥ 20 anos de idade) das Regiões Nordeste (2003, n = 75.652 e 2008, n = 79.779) e Sudeste (2003, n = 76.029 e 2008, n = 79.356) foram analisados segundo CM. Compararam-se as prevalências de CM segundo as variáveis exploratórias demográficas e de saúde no primeiro (D1) e último (D10) decil de renda familiar per capita. As análises consideraram o desenho amostral complexo. RESULTADOS: A proporção de pessoas com CM aumentou no período na Região Nordeste (61,2 para 66,9%) e Sudeste (67,9 para 73,5%). A diferença absoluta de CM, segundo D1 e D10 no período, foi de 6,4 pontos percentuais (pp) no Nordeste e 4,2 pp no Sudeste. Houve importante redução das desigualdades entre os homens; naqueles sem doenças crônicas; naqueles que tinham uma percepção positiva da sua saúde e naqueles sem plano de saúde com direito a CM. A Região Sudeste ainda apresentou redução entre aqueles com apenas uma morbidade autorreferida (8 pp) e com percepção negativa da saúde (6 pp). CONCLUSÃO: Houve aumento de CM no Brasil. Observa-se ainda persistente desigualdade entre os mais pobres e os mais ricos, maior no Nordeste do que no Sudeste. Políticas para a redução da desigualdade em saúde mais eficazes e equânimes devem ser adotadas no Brasil. .


OBJECTIVE: To analyze the socioeconomic inequalities in medical visits (MV) in the past year in Brazil. METHODS: Data from adults aged ≥ 20 years old who participated in the Brazilian National Household Surveys and living in the Northeastern (2003; n = 75,652 and 2008, n = 79,779) and Southeastern (2003; n = 76,029 and 2008; n = 79,356) regions were analyzed according to MV. We compared MVs according to demographic and health variables in the first (D1) and last (D10) per capita family income deciles. All analyses considered the complex cluster design. RESULTS: The proportion of people who had MV during this period increased in the Northeastern (from 61.2 to 66.9%) and the Southeastern (from 67.9 to 73.5%) regions. The absolute difference (AD) in the use of MV, according to D1 and D10 in this period, was equal to 6.4 percentage points (pp) in the Northeastern and 4.2 pp in the Southeastern regions. Significant reduction in inequalities was observed among men without chronic diseases, in those who had a positive perception of their health, and among those without health insurance which included MV. The Southeastern region has also showed significant reduction among those with chronic disease (8 pp) and with negative health self-perception (6 pp). CONCLUSION: The increasing number of MVs was found in Brazil. However, persistent inequalities were observed between the poorest and the richest, higher in the Northeastern than in the Southeastern region. More effective and equitable policies to reduce health inequalities should be adopted in Brazil. .


Subject(s)
Animals , Female , Humans , Mice , Adenocarcinoma/pathology , Colonic Neoplasms/pathology , Liver Neoplasms/pathology , Liver Neoplasms/secondary , Neoplasm Metastasis/pathology , Adenocarcinoma/drug therapy , Antimetabolites, Antineoplastic/therapeutic use , Antineoplastic Agents, Phytogenic/therapeutic use , Cell Adhesion , Cell Line, Tumor , Camptothecin/analogs & derivatives , Camptothecin/therapeutic use , Collagen Type II/metabolism , Colonic Neoplasms/drug therapy , Fibronectins/metabolism , Floxuridine/therapeutic use , Fluorouracil/therapeutic use , Laminin/metabolism , Liver Neoplasms/drug therapy , Mice, Nude , Models, Biological , Neoplasm Metastasis/prevention & control , Paclitaxel/therapeutic use
8.
Salud pública Méx ; 56(5): 502-510, sep.-oct. 2014. ilus, tab
Article in English | LILACS | ID: lil-733323

ABSTRACT

Objective. To estimate the annual cost of the National Cervical Cancer Screening Program (CCSP) of the Mexican Institute of Social Security (IMSS). Materials and methods. This cost analysis examined regional coverage rates reported by IMSS. We estimated the number of cytology, colposcopy, biopsy and pathology evaluations, as well as the diagnostic test and treatment costs for cervical intraepithelial neoplasia grade II and III (CIN 2/3) and cervical cancer. Diagnostic test costs were estimated using a micro-costing technique. Sensitivity analyses were performed. Results. The cost to perform 2.7 million cytology tests was nearly 38 million dollars, which represents 26.1% of the total program cost (145.4 million). False negatives account for nearly 43% of the program costs. Conclusion. The low sensitivity of the cytology test generates high rates of false negatives, which results in high institutional costs from the treatment of undetected cervical cancer cases.


Objetivo. Estimar el costo anual del Programa Nacional de Detección Oportuna de Cáncer Cervical en el Instituto Mexicano del Seguro Social (IMSS). Material y métodos. Este análisis de costos examinó las distintas coberturas por región reportadas por el IMSS. Se estimó el número de citologías, colposcopías, biopsias y evaluaciones de patología y los costos de pruebas de diagnóstico y de tratamientos por neoplasia cervical intraepitelial de grado II y III (NIC 2/3) y cáncer cervical. Los costos de las pruebas de diagnóstico se estimaron utilizando una técnica de microcosteo. Se llevó a cabo un análisis de sensibilidad. Resultados. El costo de realizar 2.7 millones de citologías fue de 38 millones de dólares, lo que representa 26.1% del costo total del programa (145.4 millones). Los falsos negativos corresponden a casi 43% de los costos del programa. Conclusiones. La baja sensibilidad de la citología genera un alto número de falsos negativos que resultan en costos elevados para la institución por el tratamiento de estos casos no detectados.


Subject(s)
Animals , Female , Rats , Collagen/metabolism , Fibronectins/metabolism , Laminin/metabolism , Liver Cirrhosis, Experimental/metabolism , Malonates/pharmacology , Antibody Specificity , Dimethylnitrosamine , Evaluation Studies as Topic , Extracellular Matrix/metabolism , Immunoenzyme Techniques , Liver Cirrhosis, Experimental/chemically induced , Rats, Sprague-Dawley
9.
Article in English | WPRIM | ID: wpr-14498

ABSTRACT

Several studies have reported that the citrus red mites Panonychus citri were an important allergen of citrus-cultivating farmers in Jeju Island. The aim of the present study was to purify and assess properties of a cysteine protease from the mites acting as a potentially pathogenic factor to citrus-cultivating farmers. A cysteine protease was purified using column chromatography of Mono Q anion exchanger and Superdex 200 HR gel filtration. It was estimated to be 46 kDa by gel filtration column chromatography and consisted of 2 polypeptides, at least. Cysteine protease inhibitors, such as trans poxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) and iodoacetic acid (IAA) totally inhibited the enzyme activities, whereas serine or metalloprotease inhibitors did not affect the activities. In addition, the purified enzyme degraded human IgG, collagen, and fibronectin, but not egg albumin. From these results, the cysteine protease of the mites might be involved in the pathogenesis such as tissue destruction and penetration instead of nutrient digestion.


Subject(s)
Animals , Chromatography, Gel , Chromatography, Ion Exchange , Collagen/metabolism , Cysteine Proteases/chemistry , Cysteine Proteinase Inhibitors/metabolism , Fibronectins/metabolism , Humans , Immunoglobulin G/metabolism , Molecular Weight , Protein Subunits/chemistry , Proteolysis , Substrate Specificity , Tetranychidae/enzymology
10.
Braz. j. microbiol ; 45(1): 263-270, 2014. tab
Article in English | LILACS | ID: lil-709487

ABSTRACT

Biofilm formation and adherence of bacteria to host tissue are one of the most important virulence factors of methicillin-resistant strains of Staphylococcus aureus (MRSA). The number of resistant strains is seriously increasing during the past years and bacteria have become resistant, not only to methicillin, but also to other commonly used antistaphylococcal antibiotics. There is a great need for discovering a novel antimicrobial agent for the treatment of staphylococcal infections. One of the most promising groups of compounds appears to be chalcones. In present study we evaluated the in vitro effect of three newly synthesized chalcones: 1,3-Bis-(2-hydroxy-phenyl)-propenone, 3-(3Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone and 3-(4-Hydroxy-phenyl)-1-(2-hydroxyphenyl)-propenone on glycocalyx production, biofilm formation and adherence to human fibronectin of clinical isolates and laboratory control strain of MRSA (ATCC 43300). Subinhibitory concentrations of the tested compounds reduced the production of glycocalyx, biofilm formation and adherence to human fibronectin of all MRSA strains. Inhibition of biofilm formation was dose dependent and the most effective was 1,3-Bis-(2-hydroxy-phenyl)-propenone. In our study we demonstrated that three newly-synthesized chalcones exhibited significant effect on adherence and biofilm formation of MRSA strains. Chalcones may be considered as promising new antimicrobial agents that can be used for prevention of staphylococcal infections or as adjunct to antibiotics in conventional therapy.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Chalcones/pharmacology , Methicillin-Resistant Staphylococcus aureus/drug effects , Anti-Bacterial Agents/chemical synthesis , Bacterial Adhesion/drug effects , Biofilms/growth & development , Chalcones/chemical synthesis , Dose-Response Relationship, Drug , Fibronectins/metabolism , Glycocalyx/metabolism , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/physiology , Structure-Activity Relationship , Staphylococcal Infections/microbiology
11.
Mem. Inst. Oswaldo Cruz ; 108(7): 825-831, 1jan. 2013.
Article in English | LILACS | ID: lil-696015

ABSTRACT

Developing thymocytes interact with thymic epithelial cells (TECs) through cell-cell interactions, TEC-derived secretory moieties and extracellular matrix (ECM)-mediated interactions. These physiological interactions are crucial for normal thymocyte differentiation, but can be disrupted in pathological situations. Indeed, there is severe thymic atrophy in animals acutely infected with Trypanosoma cruzi due to CD4+CD8+ thymocyte depletion secondary to caspase-mediated apoptosis, together with changes in ECM deposition and thymocyte migration. We studied an in vitro model of TEC infection by T. cruzi and found that infected TEC cultures show a reduced number of cells, which was likely associated with decreased proliferative capacity, but not with increased cell death, as demonstrated by bromodeoxyuridine and annexin-V labelling. The infected TEC cultures exhibited increased expression of fibronectin (FN), laminin (LM) and type IV collagen. Importantly, treatment with FN increased the relative number of infected cells, whereas treatment with anti-FN or anti-LM antibodies resulted in lower infection rates. Consistent with these data, we observed increased thymocyte adhesion to infected TEC cultures. Overall, these results suggest that ECM molecules, particularly FN, facilitate infection of the thymic epithelium and that the consequent enhancement of ECM expression might be associated with changes in TEC-thymocyte interactions.


Subject(s)
Animals , Male , Chagas Disease/metabolism , Epithelial Cells/metabolism , Extracellular Matrix/metabolism , Fibronectins/metabolism , Thymocytes/metabolism , Thymus Gland/metabolism , Cell Adhesion/physiology , Cell Communication/physiology , Cell Movement/physiology , Disease Models, Animal , Epithelial Cells/parasitology , Mice, Inbred BALB C , Thymocytes/parasitology , Thymus Gland/cytology
12.
Yonsei Medical Journal ; : 246-252, 2013.
Article in English | WPRIM | ID: wpr-17422

ABSTRACT

PURPOSE: Cell transplantation of myelin-producing exogenous cells is being extensively explored as a means of remyelinating axons in X-linked adrenoleukodystrophy. We determined whether 3,3',5-Triiodo-L-thyronine (T3) overexpresses the ABCD2 gene in the polysialylated (PSA) form of neural cell adhesion molecule (NCAM)-positive cells and promotes cell proliferation and favors oligodendrocyte lineage differentiation. MATERIALS AND METHODS: PSA-NCAM+ cells from newborn Sprague-Dawley rats were grown for five days on uncoated dishes in defined medium with or without supplementation of basic fibroblast growth factor (bFGF) and/or T3. Then, PSA-NCAM+ spheres were prepared in single cells and transferred to polyornithine/fibronectin-coated glass coverslips for five days to determine the fate of the cells according to the supplementation of these molecules. T3 responsiveness of ABCD2 was analyzed using real-time quantitative polymerase chain reaction, the growth and fate of cells were determined using 5-bromo-2-deoxyuridine incorporation and immunocytochemistry, respectively. RESULTS: Results demonstrated that T3 induces overexpression of the ABCD2 gene in PSA-NCAM+ cells, and can enhance PSA-NCAM+ cell growth in the presence of bFGF, favoring an oligodendrocyte fate. CONCLUSION: These results may provide new insights into investigation of PSA-NCAM+ cells for therapeutic application to X-linked adrenoleukodystrophy.


Subject(s)
ATP-Binding Cassette Transporters/metabolism , Adrenoleukodystrophy/genetics , Animals , Animals, Newborn , Bromodeoxyuridine , Cell Differentiation , Fibroblast Growth Factor 2/pharmacology , Fibronectins/metabolism , Immunohistochemistry , Neural Cell Adhesion Molecules/genetics , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Sialic Acids/metabolism , Stem Cells , Thyroid Hormones/metabolism , Triiodothyronine/pharmacology
13.
Yonsei Medical Journal ; : 437-444, 2013.
Article in English | WPRIM | ID: wpr-89564

ABSTRACT

PURPOSE: The present study was designed to determine whether rapamycin could inhibit transforming growth factor beta1 (TGF-beta1)-induced fibrogenesis in primary lung fibroblasts, and whether the effect of inhibition would occur through the mammalian target of rapamycin (mTOR) and its downstream p70S6K pathway. MATERIALS AND METHODS: Primary normal human lung fibroblasts were obtained from histological normal lung tissue of 3 patients with primary spontaneous pneumothorax. Growth arrested, synchronized fibroblasts were treated with TGF-beta1 (10 ng/mL) and different concentrations of rapamycin (0.01, 0.1, 1, 10 ng/mL) for 24 h. We assessed m-TOR, p-mTOR, S6K1, p-S6K1 by Western blot analysis, detected type III collagen and fibronectin secreting by ELISA assay, and determined type III collagen and fibronectin mRNA levels by real-time PCR assay. RESULTS: Rapamycin significantly reduced TGF-beta1-induced type III collagen and fibronectin levels, as well as type III collagen and fibronectin mRNA levels. Furthermore, we also found that TGF-beta1-induced mTOR and p70S6K phosphorylation were significantly down-regulated by rapamycin. The mTOR/p70S6K pathway was activated through the TGF-beta1-mediated fibrogenic response in primary human lung fibroblasts. CONCLUSION: These results indicate that rapamycin effectively suppresses TGF-beta1-induced type III collagen and fibronectin levels in primary human lung fibroblasts partly through the mTOR/p70S6K pathway. Rapamycin has a potential value in the treatment of pulmonary fibrosis.


Subject(s)
Cells, Cultured , Collagen Type III/metabolism , Fibroblasts/drug effects , Fibronectins/metabolism , Humans , Lung/cytology , Pulmonary Fibrosis/drug therapy , Signal Transduction/drug effects , Sirolimus/pharmacology , TOR Serine-Threonine Kinases/metabolism , Transforming Growth Factor beta1/antagonists & inhibitors
14.
Arq. bras. cardiol ; 99(6): 1082-1091, dez. 2012. ilus, graf, tab
Article in Portuguese | LILACS | ID: lil-662371

ABSTRACT

FUNDAMENTO: O Imatinib é um inibidor do receptor tirosina-quinase que foi confirmada como exercendo um efeito inibidor sobre a atividade do receptor do PDGF, fator de crescimento plaquetário (PDGFRα e PDGFRβ). OBJETIVO: Investigar o efeito protetor do Imatinib na fibrose miocárdica em acetato de deoxicorticosterona (DOCA)/ratos com hipertensão induzida por sal. MÉTODOS: Sessenta ratos Sprague-Dawley machos, uninefrectomizados foram distribuídos em três grupos: ratos controles (grupo CON): grupo deoxicorticosterona (grupo DOCA); grupo deoxicorticosterona e Imatinib (grupo DOCA IMA). A Pressão Arterial Sistólica (PAS) foi medida quinzenalmente. Foi estudada a porção apical do ventrículo esquerdo. Foram empregados: coloração vermelho sirius, coloração de hematoxilina-eosina, imuno-histoquímica e ensaio de western blot. RESULTADOS: A PAS nos grupos DOCA e IMA+DOCA foi maior que no grupo CON nos dias 14 e 28. Os animais do grupo DOCA apresentaram fibrose intersticial e perivascular grave no dia 28, e as expressões de PI, PIII, tenascina-C e fibronectina foram significativamente maiores que nos grupos DOCA+IMA e CON. Quando comparados com o grupo CON, os grupos DOCA e DOCA+IMA apresentaram resposta inflamatória de tecido miocárdico e infiltração de monócitos/macrófagos de diferentes graus. As expressões proteicas do PDGF-A, PDGF-C e PDGFRα foram significativamente maiores nos grupos DOCA e DOCA+IMA que no grupo CON, mas a expressão proteica do p-PDGFRα no grupo DOCA+IMA foi menor que no DOCA. CONCLUSÃO: O Imatinib pode exercer efeitos inibitórios sobre a fibrose miocárdica em ratos com hipertensão induzida por DOCA/sal, os quais podem ser atribuídos à inibição da atividade do PDGFR-α.


BACKGROUND: Imatinib is a tyrosine kinase receptor inhibitor that has been confirmed to exert inhibitory effect on the platelet derived growth factor PDGF receptor (PDGFRα and PDGFRβ) activity. OBJECTIVE: To investigate the protective effect of imatinib on the myocardial fibrosis in deoxycorticosterone-acetate (DOCA)/salt induced hypertensive rats. METHODS: Sixty male uninephrectomized Sprague-Dawley rats were assigned to three groups: control rats (CON group); deoxycorticosterone group (DOCA group); deoxycorticosterone and imatinib group (DOCA+IMA group). Systolic blood pressure (SBP) was measured biweekly. The apical portion of the left ventricle was studied. Sirius-Red staining, Hematoxylin-Eosin staining, immunohistochemistry and Western blot assay were employed. RESULTS: SBP in the DOCA group and DOCA+IMA group was higher than that in the CON group on day 14 and 28. Animals in the DOCA group showed severe interstitial and perivascular fibrosis on day 28, and the expressions of PI, PIII, tenascin-C and fibronectin were significantly higher than those in the DOCA+IMA group and CON group. When compared with the CON group, myocardial tissue inflammatory response and monocyte/macrophage infiltration of different degrees were observed in the DOCA group and DOCA+IMA group. Protein expressions of PDGF-A, PDGF-C and PDGFRα were signiflcantly higher in the DOCA and DOCA+IMA groups than those in the CON group, but the p-PDGFRα protein expression in the DOCA+IMA group was lower than that in the DOCA group. CONCLUSION: Imatinib can exert inhibitory effects on myocardial fibrosis in DOCA/salt induced hypertensive rats, which may be attributed to the inhibition of PDGFR-α activity.


Subject(s)
Animals , Male , Rats , Benzamides/pharmacology , Endomyocardial Fibrosis/drug therapy , Piperazines/pharmacology , Protein Kinase Inhibitors/pharmacology , Pyrimidines/pharmacology , Receptor, Platelet-Derived Growth Factor alpha/antagonists & inhibitors , Blotting, Western , Benzamides/therapeutic use , Blood Pressure/drug effects , Desoxycorticosterone , Disease Models, Animal , Endomyocardial Fibrosis/pathology , Fibronectins/analysis , Fibronectins/metabolism , Fibrosis/drug therapy , Fibrosis/pathology , Hypertension/chemically induced , Hypertension/physiopathology , Nephrectomy/methods , Piperazines/therapeutic use , Protein Kinase Inhibitors/therapeutic use , Pyrimidines/therapeutic use , Rats, Sprague-Dawley , Receptor, Platelet-Derived Growth Factor alpha/metabolism , Receptor, Platelet-Derived Growth Factor beta/antagonists & inhibitors , Receptor, Platelet-Derived Growth Factor beta/metabolism , Treatment Outcome , Tenascin/analysis , Tenascin/metabolism
15.
Braz. j. med. biol. res ; 44(5): 402-410, May 2011. ilus
Article in English | LILACS | ID: lil-586506

ABSTRACT

Basic fibroblast growth factor (bFGF) regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin and from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-inducedeffects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts from tissue biopsies of patients who underwent plastic surgery for repairing hypertrophic scars. The fibroblasts were then treated with different concentrations of bFGF (ranging from 0.1 to 1000 ng/mL). The growth of hypertrophic scar fibroblasts became slower with selective inhibition of type I collagen production after exposure to bFGF. However, type III collagen expression was affected in both normal and hypertrophic scar fibroblasts. Moreover, fibronectin expression in the normal fibroblasts was up-regulated after bFGF treatment. bFGF (1000 ng/mL) also induced mitochondrial depolarization in hypertrophic scar fibroblasts (P < 0.01). The cellular ATP level decreased in hypertrophic scar fibroblasts (P < 0.05), while it increased in the normal fibroblasts following treatment with bFGF (P < 0.01). These data suggest that bFGF has differential effects and mechanisms on fibroblasts of the normal skin and hypertrophic scars, indicating that bFGF may play a role in the early phase of skin wound healing and post-burn scar formation.


Subject(s)
Humans , Cicatrix, Hypertrophic/pathology , Collagen Type I/metabolism , Collagen Type III/metabolism , /pharmacology , Fibroblasts/drug effects , Fibronectins/metabolism , Skin/cytology , Cells, Cultured , Cicatrix, Hypertrophic/metabolism , Collagen Type I/ultrastructure , Collagen Type III/ultrastructure , Fibroblasts/metabolism , Fibroblasts/ultrastructure , Fibronectins/ultrastructure , Microscopy, Electron, Transmission , Membrane Potential, Mitochondrial/drug effects , Membrane Potential, Mitochondrial/physiology , Reverse Transcriptase Polymerase Chain Reaction , Skin/metabolism , Up-Regulation , Wound Healing
16.
Yonsei Medical Journal ; : 610-615, 2011.
Article in English | WPRIM | ID: wpr-33259

ABSTRACT

PURPOSE: Deposition of polymeric IgA1 in the kidney mesangium is the hallmark of IgA nephropathy, but the molecular mechanisms of IgA-mediated mesangial responses and inflammatory injuries remain poorly understood. We hypothesize that Toll-like receptor 4 (TLR4) is involved in IgA-induced mesangial cell activation. MATERIALS AND METHODS: Mouse mesangial cells were stimulated with lipopolysaccharide (LPS) (1 microg/mL), IgA (20 microg/mL), or both, and TLR4 expression was measured by real time RT-PCR and Western blot. Intracellular responses to LPS or IgA were assessed by Western blot for ERK1/2, JNK, p38 MAP kinases (MAPKs), Ikappa-Balpha degradation and fibronectin secretion. MCP-1 secretion was assessed by ELISA. Small interfering RNA (siRNA) of TLR4 was used to confirm that the effects were caused by TLR4 activity. RESULTS: LPS- or IgA-treatment upregulated the levels of TLR4 mRNA and protein in cultured MMC at 24 h. LPS and IgA induced rapid phosphorylation of MAPKs, but degradation of Ikappa-Balpha was observed only in LPS-treated MMC. LPS, but not IgA, induced increased secretion of MCP-1 and fibronectin at 24 h or 48 h. Combined LPS and IgA treatment did not cause additional increases in TLR4 mRNA and protein levels or Ikappa-Balpha degradation, and MCP-1 and fibronectin secretions were less than with LPS alone. LPS- or IgA-induced TLR4 protein levels and MAPK activation were inhibited by transfection with TLR4 siRNA. CONCLUSION: These results indicate that the activation of MAPKs and MCP-1 secretion are mediated by TLR4, at least in part, in IgA-treated mesangial cells. TLR4 is involved in mesangial cell injury by induction of pro-inflammatory cytokines in IgA nephropathy.


Subject(s)
Animals , Chemokine CCL2/metabolism , Enzyme-Linked Immunosorbent Assay , Extracellular Signal-Regulated MAP Kinases/metabolism , Fibronectins/metabolism , Glomerulonephritis, IGA/metabolism , I-kappa B Proteins/metabolism , Mesangial Cells/metabolism , Mice , Mice, Transgenic , Phosphorylation , RNA Interference , RNA, Messenger/metabolism , Signal Transduction , Toll-Like Receptor 4/antagonists & inhibitors
17.
Rio de Janeiro; s.n; 2011. 122 p. ilus.
Thesis in Portuguese | LILACS | ID: lil-619455

ABSTRACT

A esporotricose é uma doença micótica, infecciosa e crônica, que envolve o tecido cutâneo e subcutâneo, e que pode afetar seres humanos e animais. Esta micose sempre foi atribuída a um único patógeno, o Sporothrix schenckii, um fundo termodimórfico, que cresce como levedura a 37 oC e como micélio à temperatura ambiente. No entanto, nos últimos anos, foi demonstrado que isolados identificados como S. schenckii apresentavam grande variabilidade genética, sugerindo que esta táxon consiste em um complexo de espécies. Esta doença é causada pela implantação traumática do patógeno fúngico, porém os mecanismos de invasão e disseminação deste microorganismo, bem como as moléculas envolvidas nestes processos, ainda são pouco conhecidos. Com base nessas informações, este trabalho visa identificar moléculas de superfície deste patógeno envolvidas na interação deste fungo com proteínas matriciais, bem como analisar diferenças fenotípicas entre espécies do denominado complexo Sporothrix. Foram utilizados, neste estudo, cinco isolados de Sporothrix spp., sendo três isolados clínicos, um isolado ambiental e um isolado de gato. A virulência de cada isolado foi comparada à capacidade adesiva à proteína matricial fibronectina. Foi observado que os isolados com amior capacidade infectiva eram os que apresentavam maior capacidade adesiva à fibronectina. Verificamos então a expressão de adesinas para fibronectina na superfície de cada isolado, por Western blot, e observamos que os isolados mais virulentos e com maior capacidade adesiva expressavam mais adesinas para fibronectina. Bandas reativas com o anticorpo monoclonal contra adesina gp70 (mAb P6E7) foram reveladas nos extratos de parede celular dos isolados estudados. Análises por microscopia confocal revelaram a colocalização da gp70 com a adesina para fibronectina na superfície dos isolados. Análises filogenéticas demonstraram que os isolados estudados possuíam diferenças genotípicas capazes de agrupá-los em duas espécies...


Sporotrichosis is a chronic and infectious diseases that involves the cutaneous and subcutaneous tissue, which can affect humans and animals. This mycosis has always been attributed to a single pathogen, the Sporothrix schenckii, a dimorphic fungus, that grows as yeast at 37 oC and as mycelia at room temperature. However, in recent years, some isolates identifies as S. schenckii showed considerable genetic variability, sugesting that this taxon consists of a complex of species. This disease is caused by the traumatic inoculation of the fungal pathogen, however, the molecules involved in the invasion and dissemination of this microorganism are still poorly understood. The aim of this study is to identify surface molecules involved in the interaction of this fungus with extracellular matrix proteins and to examine phenotypic differences between species in the Sporothrix complex. Five isolates were used throughout this study, three clinical isolates, an environmental and one cat isolate. The virulence of each isolate was compared to the adhesive capacity to fibronectin. We observed that the most virulent isolates exhibited the higher capacity to interact with fibronectin. The expression of adhesins for fibronectin on the surface of each isolate was verified by Western blot. This analysis showed that the most virulent isolates expressed more fibronectin adhesins than the avirulent ones. Positive bands for the monoclonal antibody raised against gp70 adhesin (mAb P6E7) were revealed in cell wall extracts of the isolates studied. Confocal microscopy confirmed the colocalization of fibronectin and mAb P6E7 on the yeast cell surface. Molecular analysis showed genotypic differences between isolates used in this study, that can cluster than them into two species, S. schenckii and S. brasiliensis. This phylogenetic analysis revealed that the avirulent isolate was S. brasiliensis and not S. schenckii as previously thought. This new data led us o determine whether...


Subject(s)
Animals , Cell Adhesion Molecules , Sporotrichosis/microbiology , Sporotrichosis/virology , Fibronectins/analysis , Fibronectins/metabolism , Molecular Epidemiology , Sporothrix/classification , Sporothrix/genetics , Sporothrix/isolation & purification , Sporothrix/pathogenicity , Genotype , Mycological Typing Techniques , Phenotype , Species Specificity
18.
Braz. j. med. biol. res ; 43(1): 36-42, Jan. 2010. tab, ilus
Article in English | LILACS | ID: lil-535640

ABSTRACT

Transforming growth factor-â1 (TGF-â1) plays an important role in the fibrogenic process in the liver. The aim of the present study was to explore the action of TGF-â1 on fibronectin expression in rat hepatic stem-like cells and the underlying mechanisms. The level of fibronectin expression was determined in hepatic stem-like cells (WB cells) before and after TGF-â1 stimulation by RT-PCR and Western blot methods. Using immunogold transmission electron microscopy and the Western blot method, we observed the result of the expression and the distribution of cAMP, phosphorylated Smad3 and Smad7 before and after TGF-â1 treatment. The levels of fibronectin expression in both mRNA and protein increased 4- to 5-fold after TGF-â1 stimulation, reaching an optimum level after 8 h and then gradually falling back. Similarly, TGF-â1 stimulation resulted in an increase of cAMP in WB cells, peaking at 8 h. After treatment with TGF-â1 for 24 h, the expression of cAMP gradually decreased. In addition, we found that TGF-â1 treatment also contributed to the increased expression and to changes in cellular distribution of phosphorylated Smad3 (translocation from the cytoplasm to the nucleus) and Smad7 (translocation from the nucleus to the cytoplasm) in WB cells. The present study demonstrates that TGF-â is involved in the fibrogenic process in hepatic stem cells through up-regulation of fibronectin expression, and the mechanisms underlying this process may be associated with the activation of cAMP and Smad pathways.


Subject(s)
Animals , Rats , Fibronectins/metabolism , Hepatocytes/metabolism , Stem Cells/metabolism , Transforming Growth Factor beta/pharmacology , Blotting, Western , Cyclic AMP/metabolism , Fibronectins/genetics , Hepatocytes/pathology , Liver Cirrhosis/etiology , Microscopy, Electron, Transmission , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/metabolism , Smad Proteins/metabolism , Stem Cells/pathology
19.
Braz. j. otorhinolaryngol. (Impr.) ; 75(6): 821-825, nov.-dez. 2009. ilus, tab
Article in English, Portuguese | LILACS | ID: lil-539377

ABSTRACT

Edema de Reinke é doença crônica da laringe na qual a camada superficial da lâmina própria é expandida por muco espesso conferindo-lhe aspecto gelatinoso. Relaciona-se ao tabagismo e acomete, preferencialmente mulheres, as quais apresentam a voz mais grave. Suas características histológicas nem sempre conseguem diferenciá-lo das demais lesões benignas da laringe, havendo necessidade de técnicas histológicas adicionais. Objetivos: Estudar a imunoexpressão da fibronectina, do colágeno IV e da laminina no edema de Reinke por meio de técnicas imunoistoquímicas. Estudo prospectivo. Material e métodos: Blocos histológicos de 60 casos cirúrgicos de edema de Reinke foram resgatados, submetidos a novos cortes e às reações imunoistoquímicas para fibronectina, laminina e colágeno IV pelo método da Avidina Biotina Peroxidase. Todos os pacientes eram fumantes e adultos, sendo 50 mulheres e 10 homens. Resultados: As análises da imunoexpressão da fibronectina, do colágeno IV e da laminina foram mais expressivas no endotélio dos vasos (68,33 por cento, 76,66 por cento, 73,33 por cento, respectivamente), e menos relevantes na membrana basal (25,0 por cento, 5,0 por cento e 3,3 por cento, respectivamente). Conclusões: No edema de Reinke, a imunoexpressão da fibronectina, da laminina e do colágeno IV na membrana basal não apresentam relevância, havendo predomínio desses anticorpos no endotélio do vasos.


Reinke's edema is chronic laryngeal disease in which the superficial layer of the lamina propria is expanded by thick mucus, giving it a gelatin aspect. The disease is directly related to smoking and more frequent in women, who end up having a lower tone of voice. Its histological characteristics cannot always distinguish it from other benign lesions of the larynx for which additional histological techniques are necessary. AIM: to study the immunoexpression of fibronectin, collagen IV and laminin in Reinke's edema by immunohistochemical technique. Prospective study. Materials and methods: histological blocks of 60 cases of surgical Reinke's edema were saved, submitted to new cross-sections and to immunohistochemical reactions for fibronectin, laminin and collagen IV by the Avidin-Biotin-Peroxidase method. Fragments of five normal vocal folds were used as control, removed during autopsy. All patients were chronic smokers and adults- 50 women and 10 men. Results: the immunoexpression of fibronectin, collagen IV and laminin was more important in the endothelium of blood vessels (68.33 percent, 76.66 percent, 73.33 percent, respectively) and less relevant in the basement membrane (25.0 percent, 5.0 percent and 3.3 percent, respectively). Conclusions: the immunoexpression of fibronectin, laminin and of collagen IV in the basal membrane of Reinke's edema was not relevant, with a predominance of these antibodies in the endothelium of blood vessels.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Collagen/metabolism , Fibronectins/metabolism , Laminin/metabolism , Laryngeal Edema/metabolism , Chronic Disease , Immunohistochemistry , Laryngeal Edema/pathology , Laryngeal Edema/surgery , Prospective Studies , Young Adult
20.
Article in English | WPRIM | ID: wpr-121050

ABSTRACT

Adhesion through microbial surface components that recognize adhesive matrix molecules is an essential step in infection for most pathogenic bacteria. In this study, we report that LigB interacts with fibronectin (Fn) through its variable region. A possible role for LigB in bacterial attachment to host cells during the course of infection is supported by the following observations: (i) binding of the variable region of LigB to Madin-Darby canine kidney (MDCK) cells in a dose-dependent manner reduces the adhesion of Leptospira, (ii) inhibition of leptospiral attachment to Fn by the variable region of LigB, and (iii) decrease in binding of the variable region of LigB to the MDCK cells in the presence of Fn. Furthermore, we found a significant reduction in binding of the variable region of LigB to Fn using small interfering RNA (siRNA). Finally, the isothermal titration calorimetric results confirmed the interaction between the variable region of LigB and Fn. This is the first report to demonstrate that LigB binds to MDCK cells. In addition, the reduction of Fn expression in the MDCK cells, by siRNA, reduced the binding of LigB. Taken together, the data from the present study showed that LigB is a Fn-binding protein of pathogenic Leptospira spp. and may play a pivotal role in Leptospira-host interaction during the initial stage of infection.


Subject(s)
Animals , Antigens, Bacterial/genetics , Cell Line , Dogs , Enzyme-Linked Immunosorbent Assay , Fibronectins/metabolism , Immunoglobulin Variable Region/genetics , Leptospira/genetics , Microscopy, Confocal , Protein Binding/genetics , Protein Structure, Tertiary , RNA, Small Interfering/genetics
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