Effects of aflibercept and bevacizumab on cell viability, cell metabolism and inflammation in hypoxic human Müller cells.

Anti-VEGF (vascular endothelial growth factor) drugs such as aflibercept (AFL) and bevacizumab (BVZ) inhibit pathological neo-angiogenesis and vascular permeability in retinal vascular diseases. As cytokines and growth factors are produced by Müller glial cells under stressful and pathological conditions, we evaluated the in vitro effect of AFL (Eylea®, 0.5 mg/mL) and BVZ (Avastin®, 0.5 mg/mL) on cell viability/metabolism, and cytokine/growth factor production by Müller cells (MIO-M1) under cobalt chloride (CoCl2)-induced hypoxia after 24h, 48h and 72h. Cell viability/metabolism were analyzed by Trypan Blue and MTT assays and cytokine/growth factors in supernatants by Luminex xMAP-based multiplex bead-based immunoassay. Cell viability increased with AFL at 48h and 72h and decreased with BVZ or hypoxia at 24h. BVZ-treated cells showed lower cell viability than AFL at all exposure times. Cell metabolism increased with AFL but decreased with BVZ (72h) and hypoxia (48h and72h). As expected, AFL and BVZ decreased VEGF levels. AFL increased PDGF-BB, IL-6 and TNF-α (24h) and BVZ increased PDGF-BB (72h). Hypoxia reduced IL-1ß, -6, -8, TNF-α and PDGF-BB at 24h, and its suppressive effect was more prominent than AFL (EGF, PDGF-BB, IL-1ß, IL-6, IL-8, and TNF-α) and BVZ (PDGF-BB and IL-6) effects. Hypoxia increased bFGF levels at 48h and 72h, even when combined with anti-VEGFs. However, the stimulatory effect of BVZ predominated over hypoxia for IL-8 and TNF-α (24h), as well as for IL-1ß (72h). Thus, AFL and BVZ exhibit distinct exposure times effects on MIO-M1 cells viability, metabolism, and cytokines/growth factors. Hypoxia and BVZ decreased MIO-M1 cell viability/metabolism, whereas AFL likely induced gliosis. Hypoxia resulted in immunosuppression, and BVZ stimulated inflammation in hypoxic MIO-M1 cells. These findings highlight the complexity of the cellular response as well as the interplay between anti-VEGF treatments and the hypoxic microenvironment.

Effects of air pollution exposure on inflammatory and endurance performance in recreationally trained cyclists adapted to traffic-related air pollution.

The aim of the present study was to analyze the effects of traffic-related air pollution (TRAP) on markers of inflammatory, neuroplasticity, and endurance performance-related parameters in recreationally trained cyclists who were adapted to TRAP during a 50-km cycling time trial (50-km cycling TT). Ten male cyclists performed a 50-km cycling TT inside an environmental chamber located in downtown Sao Paulo (Brazil), under TRAP or filtered air conditions. Blood samples were obtained before and after the 50-km cycling TT to measure markers of inflammatory [interleukin-6 (IL-6), C-reactive protein (CRP), interleukin-10 (IL-10), intercellular adhesion molecule-1 (ICAM-1)] and neuroplasticity [brain-derived neurotrophic factor (BDNF)]. Rating of perceived exertion (RPE), heart rate (HR), and power output (PO) were measured throughout the 50-km cycling TT. There were no significant differences between experimental conditions for responses of IL-6, CRP, and IL-10 (P > 0.05). When compared with exercise-induced changes in filtered air condition, TRAP provoked greater exercise-induced increase in BDNF levels (TRAP = 3.3 ± 2.4-fold change; Filtered = 1.3 ± 0.5-fold change; P = 0.04) and lower exercise-induced increase in ICAM-1 (Filtered = 1.1 ± 0.1-fold change; TRAP = 1.0 ± 0.1-fold change; P = 0.01). The endurance performance-related parameters (RPE, HR, PO, and time to complete the 50-km cycling TT) were not different between TRAP and filtered air conditions (P > 0.05). These findings suggest that the potential negative impacts of exposure to pollution on inflammatory, neuroplasticity, and performance-related parameters do not occur in recreationally trained cyclists who are adapted to TRAP.

Air pollution and high-intensity interval exercise: Implications to anti-inflammatory balance, metabolome and cardiovascular responses.

High-intensity interval exercise (HIIE) is an effective non-pharmacological tool for improving physiological responses related to health. When HIIE is performed in urban centers, however, the exerciser is exposed to traffic-related air pollution (TRAP), which is associated with metabolic, anti-inflammatory imbalance and cardiovascular diseases. This paradoxical combination has the potential for conflicting health effects. Thus, the aim of this study was to determine the effects of HIIE performed in TRAP exposure on serum cytokines, non-target metabolomics and cardiovascular parameters. Fifteen participants performed HIIE in a chamber capable to deliver filtered air (FA condition) or non-filtered air (TRAP condition) from a polluted site adjacent to the exposure chamber. Non-target blood serum metabolomics, blood serum cytokines and blood pressure analyses were collected in both FA and TRAP conditions at baseline, 10 min after exercise, and 1 h after exercise. The TRAP increased IL-6 concentration by 1.7 times 1 h after exercise (p < 0.01) and did not change the anti-inflammatory balance (IL-10/TNF-α ratio). In contrast, FA led to an increase in IL-10 and IL-10/TNF-α ratio (p < 0.01), by 2.1 and 2.3 times, respectively. The enrichment analysis showed incomplete fatty acid metabolism under the TRAP condition (p < 0.05) 10 min after exercise. There was also an overactivity of ketone body metabolism (p < 0.05) at 10 min and at 1 h after exercise with TRAP. Exercise-induced acute decrease in systolic blood pressure (SBP) was not observed at 10 min and impaired at 1 h after exercise (p < 0.05). These findings reveal that TRAP potentially attenuates health benefits often related to HIIE. For instance, the anti-inflammatory balance was impaired, accompanied by accumulation of metabolites related to energy supply and reduction to exercise-induced decrease in SBP.

Traffic-related air pollution and endurance exercise: Characterizing non-targeted serum metabolomics profiling.

Although the exposure to traffic-related air pollution (TRAP) has emerged as one of main problem worldwide to inhabitants' health in urban centers, its impact on metabolic responses during exercise is poorly understood. The aim of study was to characterize the profile of non-target serum metabolomics during prolonged exercise performed under TRAP conditions. Ten healthy men completed two 90 min constant-load cycling trials under conditions of either TRAP or filtered air. Experimental trials were performed in a chamber located on an avenue with a high volume of vehicle traffic. Blood samples were taken at 30 min, 60 min, and 90 min of exercise. Based on Nuclear Magnetic Resonance metabolomics, the non-target analysis was used to assess the metabolic profile. Twelve, 16 and 18 metabolites were identified as discriminants. These were: at 30 min of exercise, the coefficient of determination (R2) 0.98, the predictive relevance, (Q2) 0.12, and the area under the curve (AUC) 0.91. After 60 min of exercise: (R2: 0.99, Q2: 0.09, AUC: 0.94); and at 90 min of exercise (R2: 0.91, Q2: <0.01, AUC: 0.89), respectively. The discriminant metabolites were then considered for the target analysis, which demonstrated that the metabolic pathways of glycine and serine metabolism (p = 0.03) had been altered under TRAP conditions at 30 min of exercise; arginine and proline metabolism (p = 0.04) at 60 min of exercise; and glycolysis (p = 0.05) at 90 min of exercise. The present results suggest that exposure to TRAP during prolonged exercise leads to a significant change in metabolomics, characterized by a transitional pattern and lastly, impairs the glucose metabolism.

Exercising in the urban center: Inflammatory and cardiovascular effects of prolonged exercise under air pollution.

The aim of this study was to investigate, in a well-controlled experimental environment, whether air pollution from an urban center would affect inflammatory and cardiorespiratory responses during prolonged moderate exercise (i.e., 90 min). Ten healthy men performed two experimental trials under filtered and polluted air, inside an environmental chamber located in Sao Paulo downtown, Brazil. Blood samples were obtained at rest, 30, 60, and 90 min of the exercise to determine the serum cytokines concentration, while arterial pressure was recorded immediately after the exercise. The serum cytokines were not altered until 60 min of exercise for both conditions (P > 0.05). Otherwise, at 90 min of exercise, the IL-6 (P = 0.047) and vascular endothelial growth factor (VEGF) (P = 0.026) were significantly higher and IL-10 tended to decrease (P = 0.061) in polluted air condition compared to filtered air condition. In addition, both systolic (P = 0.031) and diastolic (P = 0.009) arterial pressure were higher in polluted air condition than filtered air condition. These findings demonstrate that the exercise of longer duration (i.e., 90 min), but not of shorter duration (i.e., <60 min), performed in vehicular air pollution condition results in pronounced pro-inflammatory and increased arterial pressure responses.

Occupational effect of sugarcane biomass burning on the conjunctival mucin profile of harvest workers and residents of an adjacent town - A Brazilian panel study.

Pre-harvest burning of sugarcane fields produces large amounts of air pollutants which are known to cause health problems, including ocular surface abnormalities. In this study, we evaluated the effect of biomass burning on mucus quality and mucin gene expression (MUC1, MUC5AC, MUC16) in the conjunctiva of sugarcane workers (SWs) and residents of an adjacent town (RTs). Impression cytology samples of the inferior tarsal and bulbar conjunctiva of 78 SWs and 32 RTs were collected before (T1) and immediately after (T2) a 6-month harvest period. The neutral, acid and total mucus content of goblet cells was determined by PAS and AB staining. The levels of MUC5AC, MUC1 and MUC16 mRNA in the conjunctiva were measured by real-time PCR. Compared to RTs, SWs had higher levels of bulbar acid mucus and MUC16 mRNA and tarsal MUC5AC mRNA at T2 and lower levels of neutral mucus at T1 and T2. In the SW group, MUC1 mRNA levels were higher at T2 than at T1, but the levels of neutral and acid mucus were similar. In the RT group, acid mucus decreased and neutral mucus increased in the bulbar and tarsal conjunctiva at T2. In conclusion, our findings show that sugarcane harvesting is associated with abnormalities in mucus quality and content and changes in mucin mRNA levels on the ocular surface. This may help explain the ocular inflammatory signs and symptoms observed in subjects exposed to air pollutants and high temperatures from sugarcane biomass burning.

Pleural anthracosis as an indicator of lifetime exposure to urban air pollution: An autopsy-based study in Sao Paulo.

Many studies have been conducted to evaluate the association between air pollution and adverse health effects using a wide variety of methods to assess exposure. However, the assessment of individual long-term exposure to ambient air pollution is a challenging task and has not been evaluated in a large autopsy study. Our goal was to investigate whether exposure to urban air pollution is associated to the degree of lung anthracosis, considering modifying factors such as personal habits, mobility patterns and occupational activities. We conducted a study in Sao Paulo, Brazil from February 2017 to June 2018, combining epidemiological, spatial analysis and autopsy-based approaches. Information about residential address, socio-demographic details, occupation, smoking status, time of residence in the city and time spent commuting was collected via questionnaires applied to the next-of-kin. Images of the pleura surface from upper and lower lobes were used to quantify anthracosis in the lungs. We used multiple regression models to assess the association between the amount of carbon deposits in human lungs, measured by the fraction of pleural anthracosis (FA), and potential explanatory variables. We analyzed 413 cases and our data showed that for each additional hour spent in daily commuting, the ratio FA/(1-FA) is multiplied by 1.05 (95% confidence interval: [1.02; 1.08]). The estimated coefficient for daily hours spent in traffic was not considerably affected by the inclusion of socio-demographic variables and smoking habits. We estimate a tobacco equivalent dose of 5 cigarettes per day in a city where annual PM2.5 concentration oscillates around 25 µg/m3. Pleural anthracosis is a potential index of lifetime exposure to traffic-derived air pollution.

Workers of São Paulo city, Brazil, exposed to air pollution: Assessment of genotoxicity.

Air pollution affects all major urban centers, particularly megacities with populations greater than 10 million people. Vehicular and industrial emissions are among the most important sources of air pollutants in these cities. Air pollution composition, dose, and time of exposure can cause differential effects on human health. We have evaluated the genotoxic effects of air pollution (PM2.5 and NO2) on São Paulo city workers. Fifty-seven male individuals, 28-66 years old, with occupational exposure to air pollution, participated in this study; all worked daily outdoor shifts in São Paulo. Participants were recruited from three occupations: traffic controllers (n = 18); taxi drivers (n = 21); and workers at the Forestry Institute (n = 18). These workers were classified into two groups based on their workplace locations: Downtown Group (DT): traffic controllers and taxi drivers; Outskirts of Town Group (OT): workers at the Forestry Institute. Individual samplers of air pollution (Harvard air impactor) were used to collect PM2.5 and NO2 pollutants. Genotoxicity analysis (micronucleus test) was performed on buccal mucosa epithelial cells and peripheral blood lymphocytes. PM2.5 concentrations were significantly different between the groups (DT = 32.92 µg m-3, OT = 25.77 µg m-3; p = 0.03); however, no difference was observed in NO2 concentrations. Micronucleus frequencies in both buccal mucosa (DT = 2.78%, OT = 1.16%; p < 0.0001) and in peripheral lymphocytes (DT = 1.51%, OT = 0.73%; p < 0.0001) were significantly different between the groups. We observed a direct correlation between the individual dose of PM2.5 and micronucleus frequency in the buccal mucosa (p = 0.0021). Our results indicate that workers in the most urban areas of São Paulo are exposed to higher concentrations of PM2.5 and showed higher micronucleus frequencies in both buccal mucosa and lymphocytes.

Exercising in Air Pollution: The Cleanest versus Dirtiest Cities Challenge.

Background: Aerobic exercise is recommended to improve health. However, the increased ventilation might increase the doses of inhaled air pollutants, negating the health benefits in highly polluted areas. Our objective was to estimate the inhaled dose of air pollutants during two simulated exercise sessions at cleanest and dirtiest cities reported by World Health Organization (WHO) considering air quality. Methods: Minute ventilation data were extracted from laboratory-based exercise of 116 incremental running tests and used to calculate total ventilation of a hypothetical 30-min moderate continuous exercise routine. Afterwards, total ventilation values were combined with particulate matter (PM) data reported by the WHO for the 10 cleanest and 10 dirtiest cities, to calculate inhaled doses and the relative risk of all-cause mortality by exercising in different air pollution concentrations. Findings: The dirtiest cities are located at less developed countries compared to cleanest cities. The inhaled dose of PM2.5 and PM10 were significantly higher in the dirtiest cities compared to the cleanest cities at rest and exercise, and significantly higher during exercise compared to the rest at dirtiest cities. The relative risk of all-cause mortality analysis showed that, while exercise in the cleanest cities improved health benefits throughout up to 90 min, there were no further health benefits after 15 min of exercise in the dirtiest cities, and the air pollution health risks surpassed the exercise benefits after 75 min. Interpretation: Our findings suggest that a traditional 30-min of moderate aerobic exercise session might induce inhalation of high levels of pollutants when performed at dirtiest cities. Considering several adverse health effects from air pollutants inhalation, so the results suggest that the air pollution levels of the cities should be taken into account for physical exercise recommendations.

Cellular stress response in human Müller cells (MIO-M1) after bevacizumab treatment.

Bevacizumab, an anti-vascular endothelial growth factor (VEGF) agent, is widely used in the treatment of retinal vascular diseases. However, due to the essential role Müller cell derived-VEGF plays in the maintenance of retinal neurons and glial cells, cell viability is likely to be affected by VEGF inhibition. We therefore evaluated the effect of bevacizumab-induced VEGF inhibition on Müller cells (MIO-M1) in vitro. MIO-M1 cells were cultured for 12 or 24 h in media containing bevacizumab at 0.25 or 0.5 mg/mL. Controls were cultured in medium only. Cell viability was determined with the trypan blue exclusion test and MTT assay. Caspase-3, beclin-1, glial fibrillary acidic protein (GFAP) and vimentin content were quantified by immunohistochemistry. Gene expression was evaluated by real-time quantitative PCR. Treatment with bevacizumab did not reduce MIO-M1 cell viability, but increased metabolic activity at 24 h (0.5 mg/mL) and induced apoptosis and autophagy, as shown by the increased caspase-3 levels at 12 h (0.25 and 0.5 mg/mL) and the increased beclin levels at 24 h (0.5 mg/mL). Caspase-3 mRNA was upregulated at 12 h and downregulated at 24 h in cells treated with bevacizumab at 0.25 mg/mL. Bevacizumab treatment was also associated with structural protein abnormalities, with decreased GFAP and vimentin content and upregulated GFAP and vimentin mRNA expression. Although bevacizumab did not significantly affect MIO-M1 cell viability, it led to metabolic and molecular changes (apoptosis, autophagy and structural abnormalities) suggestive of significant cellular toxicity.

The genetics of human running: ACTN3 polymorphism as an evolutionary tool improving the energy economy during locomotion.

BACKGROUND: Covering long distances was an important trait to human evolution and continues to be highlighted for health and athletic status. This ability is benefitted by a low cost of locomotion (CoL), meaning that the individuals who are able to expend less energy would be able to cover longer distances. The CoL has been shown to be influenced by distinct and even 'opposite' factors, such as physiological and muscular characteristics, which are genetically inherited. In this way, DNA alterations could be important determinants of the characteristics associated with the CoL. A polymorphism in the ACTN3 gene (R577X) has been related to physical performance, associating the X allele with endurance and the R allele with strength/power abilities. AIM: To investigate the influence of ACTN3 genotypes on the CoL. SUBJECTS AND METHODS: One hundred and fifty healthy male individuals performed two constant speed tests (at 10 and 12 km/h) to determine the CoL. RESULTS: Interestingly, the results showed that heterozygous individuals (RX genotype) presented significantly lower CoL compared to RR and XX individuals. CONCLUSIONS: It is argued that RX genotype might generate an intermediate strength-to-endurance phenotype, leading to a better phenotypic profile for energy economy during running and, consequently, for long-term locomotion.

Influence of ACTN3 R577X polymorphism on ventilatory thresholds related to endurance performance.

The purpose of this study was to verify the association between ACTN3 polymorphism and physiological parameters related to endurance performance. A total of 150 healthy male volunteers performed a maximal incremental running test to determine the speeds corresponding to ventilatory threshold (VT) and respiratory compensation point (RCP). Participants were genotyped and divided into terciles based on the analysed variables. Genotype frequencies were compared through χ(2) test between lower and higher terciles, with the lowest or highest values of each analysed variable. ACTN3 XX genotype was over-represented in higher tercile for VT and RCP. Odds ratio also showed significantly higher chances of XX individuals to be in higher tercile compared to RR (7.3) and RR + RX (3.5) for VT and compared to RR genotype (8.1) and RR + RX (3.4) for RCP. Thus, XX individuals could attain the VT and RCP at higher speeds, suggesting that they are able to sustain higher running speeds in lower exercise intensity domains. It could result in higher lipid acids oxidation, saving muscle glycogen and delaying the fatigue during prolonged exercises, which could be the advantage mechanism of this genotype to endurance performance.

Lacrimal Cytokines Assessment in Subjects Exposed to Different Levels of Ambient Air Pollution in a Large Metropolitan Area.

BACKGROUND: Air pollution is one of the most environmental health concerns in the world and has serious impact on human health, particularly in the mucous membranes of the respiratory tract and eyes. However, ocular hazardous effects to air pollutants are scarcely found in the literature. DESIGN: Panel study to evaluate the effect of different levels of ambient air pollution on lacrimal film cytokine levels of outdoor workers from a large metropolitan area. METHODS: Thirty healthy male workers, among them nineteen professionals who work on streets (taxi drivers and traffic controllers, high pollutants exposure, Group 1) and eleven workers of a Forest Institute (Group 2, lower pollutants exposure compared to group 1) were evaluated twice, 15 days apart. Exposure to ambient PM2.5 (particulate matter equal or smaller than 2.5 µm) was 24 hour individually collected and the collection of tears was performed to measure interleukins (IL) 2, 4, 5 and 10 and interferon gamma (IFN-γ) levels. Data from both groups were compared using Student's t test or Mann- Whitney test for cytokines. Individual PM2.5 levels were categorized in tertiles (lower, middle and upper) and compared using one-way ANOVA. Relationship between PM2.5 and cytokine levels was evaluated using generalized estimating equations (GEE). RESULTS: PM2.5 levels in the three categories differed significantly (lower: ≤22 µg/m3; middle: 23-37.5 µg/m3; upper: >37.5 µg/m3; p<0.001). The subjects from the two groups were distributed unevenly in the lower category (Group 1 = 8%; Group 2 = 92%), the middle category (Group 1 = 89%; Group 2 = 11%) and the upper category (Group 1 = 100%). A significant relationship was found between IL-5 and IL-10 and PM2.5 levels of the group 1, with an average decrease of 1.65 pg/mL of IL-5 level and of 0.78 pg/mL of IL-10 level in tear samples for each increment of 50 µg/m3 of PM2.5 (p = 0.01 and p = 0.003, respectively). CONCLUSION: High levels of PM2.5 exposure is associated with decrease of IL-5 and IL-10 levels suggesting a possible modulatory action of ambient air pollution on ocular surface immune response.

Is the COL5A1 rs12722 gene polymorphism associated with running economy?

The COL5A1 rs12722 polymorphism is considered to be a novel genetic marker for endurance running performance. It has been postulated that COL5A1 rs12722 may influence the elasticity of tendons and the energetic cost of running. To date, there are no experimental data in the literature supporting the relationship between range of motion, running economy, and the COL5A1 rs12722 gene polymorphism. Therefore, the main purpose of the current study was to analyze the influence of the COL5A1rs12722 polymorphism on running economy and range of motion. One hundred and fifty (n = 150) physically active young men performed the following tests: a) a maximal incremental treadmill test, b) two constant-speed running tests (10 km · h(-1)) and 12 km · h(-1)) to determine the running economy, and c) a sit-and-reach test to determine the range of motion. All of the subjects were genotyped for the COL5A1 rs12722 single-nucleotide polymorphism. The genotype frequencies were TT = 27.9%, CT = 55.8%, and CC = 16.3%. There were no significant differences between COL5A1 genotypes for running economy measured at 10 km · h(-1) (p = 0.232) and 12 km · h(-1) (p = 0.259). Similarly, there were no significant differences between COL5A1 genotypes for range of motion (p = 0.337). These findings suggest that the previous relationship reported between COL5A1 rs12722 genotypes and running endurance performance might not be mediated by the energetic cost of running.

Bevacizumab reduces neurocan content and gene expression in newborn rat retina in vitro.

PURPOSE: Extracellular matrix (ECM) and cellular membrane proteoglycans (PGs) play important roles in neural differentiation and cell adhesion. Vascular endothelial growth factor, an important signal protein in vascular and retinal neural cell development, is retained in the ECM due to its high affinity for PG. Bevacizumab, an anti-VEGF agent, has been extensively used for treating retinal diseases in adult and newborn patients, although its effect on the developing retina remains largely unknown. The purpose of this study was to investigate the effect of bevacizumab on neurocan, phosphacan, and syndecan-3 PG levels in newborn rat retina. METHODS: Retinal explants of sixty 2-day-old Lister hooded rats were obtained after eye enucleation and maintained in culture media with or without bevacizumab for 48 hours. Immunohistochemical staining was assessed against neurocan, phosphacan, and syndecan-3. Proteoglycan content was quantified based on the intensity of immunohistochemical labeling. Gene expressions were quantified by real-time reverse-transcription polymerase chain reaction. The results from the treatment and control groups were compared. RESULTS: No significant difference in the staining intensity and mRNA expression of phosphacan and syndecan-3 was observed between the groups. However, a significant decrease in neurocan content and mRNA expression was observed in bevacizumab-treated retinal explants compared with controls. CONCLUSIONS: Bevacizumab did not affect phosphacan and syndecan-3 levels but decreased neurocan content and gene expression. Therefore, it may interfere with early postnatal retinal cell differentiation. Although further studies are necessary to confirm our findings, we suggest anti-VEGF agents be used with caution in developing retinal tissue.

Changes in cardiac heparan sulfate proteoglycan expression and streptozotocin-induced diastolic dysfunction in rats.

BACKGROUND: Changes in the proteoglycans glypican and syndecan-4 have been reported in several pathological conditions, but little is known about their expression in the heart during diabetes. The aim of this study was to investigate in vivo heart function changes and alterations in mRNA expression and protein levels of glypican-1 and syndecan-4 in cardiac and skeletal muscles during streptozotocin (STZ)-induced diabetes. METHODS: Diabetes was induced in male Wistar rats by STZ administration. The rats were assigned to one of the following groups: control (sham injection), after 24 hours, 10 days, or 30 days of STZ administration. Echocardiography was performed in the control and STZ 10-day groups. Western and Northern blots were used to quantify protein and mRNA levels in all groups. Immunohistochemistry was performed in the control and 30-day groups to correlate the observed mRNA changes to the protein expression. RESULTS: In vivo cardiac functional analysis performed using echocardiography in the 10-day group showed diastolic dysfunction with alterations in the peak velocity of early (E) diastolic filling and isovolumic relaxation time (IVRT) indices. These functional alterations observed in the STZ 10-day group correlated with the concomitant increase in syndecan-4 and glypican-1 protein expression. Cardiac glypican-1 mRNA and skeletal syndecan-4 mRNA and protein levels increased in the STZ 30-day group. On the other hand, the amount of glypican in skeletal muscle was lower than that in the control group. The same results were obtained from immunohistochemistry analysis. CONCLUSION: Our data suggest that membrane proteoglycans participate in the sequence of events triggered by diabetes and inflicted on cardiac and skeletal muscles.

Gene expression of extracellular matrix proteoglycans in human cyclosporin-induced gingival overgrowth.

BACKGROUND: Gingival overgrowth is one of the side effects associated with the systemic use of cyclosporin A (CsA). In vitro studies on the extracellular matrix of gingival tissues have demonstrated an altered composition, particularly an accumulation of proteoglycans and collagen. We investigated the gene expression of extracellular matrix proteoglycans in CsA-induced gingival tissue alterations. METHODS: mRNA expression of the proteoglycans perlecan, decorin, biglycan, and versican was analyzed by reverse transcription polymerase chain reaction (RT-PCR) in gingival samples obtained from 12 individuals, six with CsA-induced gingival overgrowth (CsA group) and six with a normal gingiva (control group). The RT-PCR products were subjected to 1% agarose gel electrophoresis containing ethidium bromide and analyzed qualitatively and semiquantitatively by densitometry. Density values were normalized by determining the expression of the housekeeping gene beta-actin in the same sample. Groups were compared by the Student's t test. RESULTS: Perlecan expression showed a marked increase (54%) in the CsA group compared to the control group (P < 0.01), while no significant differences were observed for the other proteoglycans. CONCLUSION: CsA-induced gingival overgrowth seems to be associated with increased expression of perlecan, a typical basement membrane proteoglycan, but not decorin, biglycan, or versican.

Distribuição comparativa dos glicosaminoglicanos em artérias e veias de diferentes mamíferos / Comparative distribution of glycosaminoglycans in arteries and veins of different mammals

Foi realizada análise comparativa sobre a distribuição dos glicosaminoglicanos de artérias e veias em ratos, cachorros e humanos. Os nossos resultados demonstraram que dermatam sulfato foi o principal glicosaminoglicano encontrado tanto para as artérias quanto para as veias estudadas. Entretanto, a proporção de dermatam sulfato foi maior nas veias do que nas artérias nas três espécies analisadas. Este aumento pode estar associado às diferenças estruturais e funcionais encontradas na parede destes dois tipos de vasos sangüíneos (nas veias a pressão sangüínea é significativamente mais baixa). Além disso, a quantidade total dos glicosaminoglicanos foi maior nas artérias do que nas veias, sendo as maiores concentrações encontradas nas aortas independentemente da espécie animal estudada. Estes achados abrem perspectiva para o melhor conhecimento das alterações das macromoléculas que possam estar relacionadas ao processo degenerativo vascular, especialmente nas transformações estruturais que as veias safenas sofrem, quando empregadas como enxertos na revascularização do miocárdio