Genes, calcium and modifying factors in hypertrophic cardiomyopathy

1. Familial hypertrophic cardiomyopathy (FHC) is a primary disorder of the myocardium characterized by remarkable diversity in clinical presentations, ranging from no symptoms to severe heart failure and sudden cardiac death. 2. Over the past 15 years, at least 11 genes have been identified, defects of which cause FHC. Most of these genes encode proteins that comprise the basic contractile unit of the heart (i.e. the sarcomere). 3. Genetic studies are now beginning to have a major impact on the diagnosis in FHC, as well as in guiding treatment and preventative strategies. Although much is known about which genes cause disease, relatively little is known about the molecular steps leading from the gene defect to the clinical phenotype and what factors modify the expression of the mutant genes. 4. Concurrent studies in cell culture and animal models of FHC are now beginning to shed light on the signalling pathways involved in FHC and the role of both environmental and genetic modifying factors. Calcium dysregulation appears to be important in the pathogenesis of FHC. 5. Understanding these basic molecular mechanisms will ultimately improve our knowledge of the basic biology of heart muscle function and will therefore provide new avenues for diagnosis and treatment not only for FHC, but also for a range of human cardiovascular diseases.     

The effect of methadone and buprenorphine on human placental aromatase

Methadone and buprenorphine (BUP) are used for treatment of the pregnant opiate addict. CYP19/aromatase is the major placental enzyme responsible for the metabolism of methadone to 2-ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and BUP to norbuprenorphine (norBUP). The aim of this investigation was to determine the effects of methadone and BUP on the activity of placental microsomal aromatase in the conversion of its endogenous substrates testosterone to 17beta-estradiol (E(2)) and 16alpha-hydroxytestosterone (16-OHT) to estriol (E(3)). The conversion of testosterone and 16-OHT by human placental microsomes exhibited saturation kinetics, and the apparent K(m) values were 0.2 +/- 1 and 6 +/- 3 microM, respectively. V(max) values for E(2) and E(3) formation were 70 +/- 16 and 28 +/- 10 pmol/mg proteinmin, respectively. Also, data obtained revealed that methadone and BUP are competitive inhibitors of testosterone conversion to E(2) and 16-OHT to E(3). The K(i) for methadone inhibition of E(2) and E(3) formation were 393 +/-144 and 53 +/- 28 microM, respectively, and for BUP the K(i) was 36 +/- 9 and 6 +/- 1 microM. The higher potency of the two opiates and their metabolites in inhibiting E(3) formation is in agreement with the lower affinity of 16-OHT than testosterone to aromatase. Moreover, the metabolites EDDP and norBUP were weaker inhibitors of aromatase than their parent compounds. The determined inhibition constants of methadone and BUP for E(3) formation by a cDNA-expressed CYP19 preparation were similar to those for placental microsomes. Therefore, data reported here suggest that methadone, BUP, and their metabolites are inhibitors of androgen aromatization in the placental biosynthesis of estrogens.     

Effect of pyridoxylidene aminoguanidine on human diploid cells B-HEF-2: in vitro cytotoxicity test and cytogenetic analysis.

BACKGROUND: Pyridoxylidene aminoguanidine is an appropriate inhibitor of protein glycation, respectively formation of advanced glycation products, which are connected with mechanism of pathogenesis in chronic diabetic complications. Moreover, it was found that in comparison with aminoguanidine, pyridoxylidene aminoguanidine does not influence the level of vitamin B6 in liver and kidneys in vivo. The aim of this study was to test cytotoxic effect of pyridoxylidene aminoguanidine in vitro, in regard to its potential use as inhibitor of advance protein glycation in diabetic patients. METHODS: The potential genotoxic activity of pyridoxylidene aminoguanidine in vitro was assessed by the micronucleus test and the karyological analysis. The direct contact method using diploid human cell line B-HEF-2 was performed to evaluate cytotoxicity. The concentrations of 5 x 10(-3), 2.5 x 10(-3) and 1 x 10(-3) ml/l were used in all tests. RESULTS: Microscopic analysis did not proved any changes in morphology of exposed fibroblasts. The inhibitive effect of pyridoxylidene aminoguanidine was increased with rising concentration. The proliferative activity of exposed cells to concentrations of 1 x 10(-3), 2.5 x 10(-3), 5 x 10(-3) mol/l was inhibited approximately by 30%, 60% and 80%, respectively. The frequency of micronuclei and rate of numerical or structural aberrations was not increased. CONCLUSION: Obtained results confirmed that pyridoxylidene aminoguanidine in selected concentrations has an inhibitive effect on the proliferation activity of exposed cells, but did not develop any cytotoxic effect on B-HEF-2 cells.     

Mortality among Guarani Indians in Southeastern and Southern Brazil

Worldwide, indigenous peoples display a high burden of disease, expressed by profound health inequalities in comparison to non-indigenous populations. This study describes mortality patterns among the Guarani in Southern and Southeastern Brazil, with a focus on health inequalities. The Guarani population structure is indicative of high birth and death rates, low median age and low life expectancy at birth. The crude mortality rate (crude MR = 5.0/1,000) was similar to the Brazilian national rate, but the under-five MR (44.5/1,000) and the infant mortality rate (29.6/1,000) were twice the corresponding MR in the South and Southeast of Brazil. The proportion of post-neonatal infant deaths was 83.3%, 2.4 times higher than general population. The proportions of ill-defined (15.8%) and preventable causes (51.6%) were high. The principal causes of death were respiratory (40.6%) and infectious and parasitic diseases (18.8%), suggesting precarious living conditions and deficient health services. There is a need for greater investment in primary care and interventions in social determinants of health in order to reduce the health inequalities.     

Estimating the potential of taxes on sugar-sweetened beverages to reduce consumption and generate revenue

Objective

Beverage taxes came into light with increasing concerns about obesity, particularly among youth. Sugar-sweetened beverages have become a target of anti-obesity initiatives with increasing evidence of their link to obesity. Our paper offers a method for estimating revenues from an excise tax on sugar-sweetened beverages that governments of various levels could direct towards obesity prevention.

Model

We construct a model projecting beverage consumption and tax revenues based on best available data on regional beverage consumption, historic trends and recent estimates of the price elasticity of sugar-sweetened beverage demand.

Results

The public health impact of beverage taxes could be substantial. An estimated 24% reduction in sugar-sweetened beverage consumption from a penny-per-ounce sugar-sweetened beverage tax could reduce daily per capita caloric intake from sugar-sweetened beverages from the current 190-200 cal to 145-150 cal, if there is no substitution to other caloric beverages or food. A national penny-per-ounce tax on sugar-sweetened beverages could generate new tax revenue of $79 billion over 2010-2015.

Conclusion

A modest tax on sugar-sweetened beverages could both raise significant revenues and improve public health by reducing obesity. To the extent that at least some of the tax revenues get invested in obesity prevention programs, the public health benefits could be even more pronounced.     

Biochemical and biological characterization of the venoms of Bothriopsis bilineata and Bothriopsis taeniata (Serpentes: Viperidae).

Snake venom is a complex mixture containing diverse protein components with different structures and functions that are used for prey immobilization and death. Snake venoms from the family Viperidae cause pronounced local and systemic effects, such as pain, edema, hemorrhage and necrosis. Here, we investigated the enzymatic and biological activities of venoms from two Amazonian snakes, Bothriopsis bilineata and Bothriopsis taeniata. Both venoms presented high enzymatic activities for proteases kallikrein, thrombin and plasmin, low levels of trypsin, cathepsin C and leucine aminopeptidase activities, while lacked acetylcholinesterase activity. B. taeniata and B. bilineata crude venoms caused inflammation inducing neutrophil recruitment into peritoneal cavity of mice 4h after injection. Neutrophil recruitment induced by B. taeniata venom was accompanied by hemorrhage. EDTA treatment profoundly impaired neutrophil recruitment, suggesting the involvement of a metalloproteinase on venoms-induced neutrophil recruitment. Pretreatment with dexamethasone and zileuton, a 5-lipoxygenase inhibitor, significantly reduced neutrophil migration, but indomethacin and montelukast, a cysteinyl leukotriene receptor antagonist, had no effect, suggesting the involvement of lipoxygenase-derived metabolites, probably LTB(4). Together, these results show that B. bilineata and B. taeniata venoms induce a marked inflammatory reaction, with leukocyte recruitment, and hemorrhage, which parallels to a high proteolytic activity found in these venoms.     

Extracellular matrix-mediated control of aortic smooth muscle cell growth and migration by a combination of ascorbic acid, lysine, proline, and catechins

Extracellular matrix (ECM) function and structure are severely compromised at atherosclerotic lesion sites, contributing to initiation and progression of the disease. This study investigated whether ECM biological properties would be beneficially affected by exposure to nutrients essential for collagen synthesis and posttranslational modification. Confluent layers of human aortic smooth muscle cells (SMC) grown on collagen substrate were cultured in the presence of the tested compounds for 7 to 10 days. Pretreated cells were removed from the ECM surface by differential treatment and replaced with secondary innocent SMC cultures. Secondary SMC growth rate and invasiveness were assayed in standard growth medium. ECM protein composition was assayed immunochemically. ECM produced in the presence of ascorbic acid reduced SMC proliferation in a dose-dependent manner. Plant-derived phenolic extracts expressed different degrees of SMC growth inhibition when present during ECM production. A combination of selected nutrients had a greater effect than did individual components. The ECM deposited by SMC in the presence of ascorbate, lysine, proline, and green tea catechins inhibited SMC migration rate up to 70%. The ECM produced under conditions of chronic essential nutrient deficiency can support proatherosclerotic SMC behavior. A combination of selected nutrients can counteract these adverse effects stronger than individual components.     

Residual monomer of autopolymerized acrylic resin according to different manipulation and polishing methods. An in situ evaluation

OBJECTIVE: To test the hypothesis that no difference exists in the in situ levels of residual monomer of an autopolymerized acrylic resin, irrespective of the method of manipulation and polishing. MATERIALS AND METHODS: Forty volunteers received the test specimens. The residual monomer was evaluated by means of gas chromatography using methylethylketone as extraction solvent. The samples of autopolymerized acrylic resin were submitted to two methods of manipulation--mass and addition--as well as to two types of polishing-mechanical and chemical. Four test groups were determined according to manipulation and polishing techniques: mass-mechanical; mass-chemical; addition-mechanical; addition-chemical. The data obtained were analyzed by the Student's t-test for paired samples as well as by analysis of variance. RESULTS: Residual methylmethacrylate was verified in high concentrations in the beginning of the testing as well as 24 hours after the test specimens had been worn. Mechanical polishing was associated with lower levels of residual monomer. The mass-mechanical group showed the lowest values. Mechanical polishing was associated with the lowest levels of residual monomer, and the lowest level of all occurred in mass-mechanical group. CONCLUSIONS: The hypothesis is rejected. Variations of residual polymer were associated with variations in the manipulation and polishing, but high concentrations of residual monomer were present in all groups. These concentrations were reduced after 24 hours.     

Influence of light-polymerization modes on the degree of conversion and mechanical properties of resin composites: a comparative analysis between a hybrid and a nanofilled composite

This study analyzed the influence of the light polymerization mode on the degree of conversion (DC) and mechanical properties of two resin composites: a hybrid (Filtek P60) and a nanofilled composite (Filtek Supreme). The composites were light activated by three light polymerization modes (Standard-S: 650 mW/cm2 for 30 seconds; High intensity-H: 1000 mW/cm2 for 20 seconds and Gradual-G: 100 up to 1000 mW/cm2 for 10 seconds + 1000 mW/cm2 for 10 seconds). The DC (%) was measured by FT-Raman spectroscopy. Flexural strength and flexural modulus were obtained from bar-shaped specimens (1 x 2 x 10 mm) submitted to the three-point bending test. Microhardness was evaluated by Knoop indentation (KHN). Data were analyzed by ANOVA and Student-Newman-Keuls multiple range test and linear regression analysis. The results showed the following DC: H > S > G (p < 0.0001) and hybrid > nanofilled (p < 0.005). Correlation was found between DC and the radiant exposure (R2 = 0.92). With respect to mechanical properties, only KHN was significantly influenced by the light polymerization mode, as follow: H > S = G (p < 0.0001). The hybrid composite presented higher flexural strength and flexural modulus than the nanofilled composite (p < 0.0001). No significant difference was found in KHN between thetwo composites (p = 0.1605). The results suggest that nanofilled composites may present a lower degree of conversion and reduced mechanical properties compared to hybrid composites.