Simplified preparation of human arterial sections for PCR analysis of Chlamydia pneumoniae and human DNA

AIMS: To investigate multiple techniques for the preparation of solid tissue for polymerase chain reaction (PCR) analysis, and to identify the most simple techniques for routine use in the laboratory. METHODS: Techniques for the preparation of arterial tissue samples including homogenisation, ultrafiltration, and treatments involving proteinase K, Gene Clean, lectin, and Fe3+ specific chelators were evaluated using the PCR to amplify both Chlamydia pneumoniae and human DNA. RESULTS: Treatment with either Gene-Clean or lectin and the Fe3+ specific chelator deferoxamine mesylate removed PCR inhibitors from tissue homogenates. Homogenisation followed by GeneClean treatment resulted in the amplification of C pneumoniae DNA from within a section of atherosclerotic carotid artery, implying that C pneumoniae elementary bodies had been disrupted. In eight further clinical samples from patients not known to have C pneumoniae infection, human DNA was amplified and no cross contamination was observed between samples. These samples contained no evidence of C pneumoniae by PCR. CONCLUSIONS: A simple preparation of solid tissue for PCR analysis, involving homogenisation followed by GeneClean treatment has been developed, and is effective for the amplification of both C pneumoniae and human DNA.

A comparative study of the effects of hypoxia upon isolated arterial muscle from the rat

Contractile response of rat aorta, mesenteric artery and femoral artery to noradrenaline and potassium chloride were studied under standard and hypoxic conditions and the effect of hypoxia was dependent upon both the vessel and the stimulant. Hypoxia had less effect upon contractions to potassium chloride than those to noradrenaline. The effects of hypoxia on potassium chloride induced responses in different vessels were relatively similar although responses to noradrenaline were vessel dependent. Noradrenaline induced contractions of the femoral artery were most affected by hypoxia whilst those of the mesenteric artery were least affected. Hypoxia changed the well maintained response of the femoral artery to noradrenaline to a transient form; this effect of hypoxia was not evident in the aorta or the mesenteric artery. The aorta and mesenteric artery contracted in calcium free EGTA PSS suggesting that these vessels displayed a release component. Hypoxia reduced the magnitude of this component. The effects of verapamil on noradrenaline and potassium chloride induced responses were investigated and were found to be different to those of hypoxia. Verapamil exerted a greater effect on contractions to potassium chloride than on those to noradrenaline. The effects of hypoxia on 45calcium flux were also vessel dependent. In the mesenteric and femoral arteries hypoxia increased basal 45calcium accumulation. However, the magnitude of noradrenaline stimulated 45calcium accumulation was reduced in the femoral artery and aorta but was unchanged in the mesenteric artery. The effects of hypoxia on 45calcium accumulation were similar to verapamil only in the aorta. The results provide evidence that the effects of hypoxia may arise from alterations in calcium mobilisation processes and that differences between vessels in these processes accounts for the heterogeneity between vessels in their response to hypoxia.

Inhibition of pyruvate kinase M2 by reactive oxygen species contributes to the development of pulmonary arterial hypertension

Aims: Pulmonary arterial hypertension [1] is a proliferative disorder associated with enhanced proliferation and suppressed apoptosis of pulmonary artery smooth muscle cells (PASMCs). Reactive oxygen species (ROS) is implicated in the development of PAH and regulates the vascular tone and functions. However, which cellular signaling mechanisms are triggered by ROS in PAH is still unknown. Hence, here we wished to characterize the signaling mechanisms triggered by ROS. Methods and Results: By Western blots, we showed that increased intracellular ROS caused inhibition of the glycolytic pyruvate kinase M2 (PKM2) activity through promoting the phosphorylation of PKM2. Monocrotaline (MCT)-induced rats developed severe PAH and right ventricular hypertrophy, with a significant increase in the P-PKM2 and decrease in pyruvate kinase activity which could be attenuated with the treatments of PKM2 activators, FBP and l-serine. The antioxidant NAC, apocynin and MnTBAP had the similar protective effects in the development of PAH. In vitro assays confirmed that inhibition of PKM2 activity could modulate the flux of glycolytic intermediates in support of cell proliferation through the increased pentose phosphate pathway (PPP). Increased ROS and decreased PKM2 activity also promoted the Cav1.2 expression and intracellular calcium. Conclusion: Our data provide new evidence that PKM2 makes a critical regulatory contribution to the PAHs for the first time. Decreased pyruvate kinase M2 activity confers additional advantages to rat PASMCs by allowing them to sustain anti-oxidant responses and thereby support cell survival in PAH. It may become a novel treatment strategy in PAH by using of PKM2 activators.

Localized retinal nerve fiber layer defects and arterial hypertension:insights into pathophysiology and perhaps an eye for detail?

Retinal microcirculatory changes are useful markers in patients suffering from hypertension and other cardiovascular disorders. Conversely, localized retinal nerve fiber layer defects (RNFLDs) are less frequently explored in association with hypertension.

On arterial contractility, energetic requirements of the cardiovascular system, and the formation of arterial sclerosis

The aorta has been viewed as a passive distribution manifold for blood whose elasticity allows it to store blood during cardiac ejection (systole), and release it during relaxation (diastole). This capacitance, or compliance, lowers peak cardiac work input and maintains peripheral sanguine irrigation throughout the cardiac cycle. The compliance of the human and canine circulatory systems have been described either as constant throughout the cycle (Toy et al. 1985) or as some inverse function of pressure (Li et al. 1990, Cappelo et al. 1995). This work shows that a compliance value that is higher during systole than diastole (equivalent to a direct function of pressure) leads to a reduction in the energetic input to the cardiovascular system (CV), even when accounting for the energy required to change compliance. This conclusion is obtained numerically, based on a 3-element lumped-parameter model of the CV, then demonstrated in a physical model built for the purpose. It is then shown, based on the numerical and physical models, on analytical considerations of elastic tubes, and on the analysis of arterial volume as a function of pressure measured in vivo (Armentano et al. 1995), that the mechanical effects of a presupposed arterial contraction are consistent with those of energetically beneficial changes in compliance during the cardiac cycle. Although the amount of energy potentially saved with rhythmically contracting arteries is small (mean 0.55% for the cases studied) the importance of the phenomenon lies in its possible relation to another function of the arterial smooth muscle (ASM): synthesis of wall matrix macromolecules. It is speculated that a reduction in the rate of collagen synthesis by the ASM is implicated in the formation of arteriosclerosis. ^

Fluid dynamics and platelet deposition in a model arterial stenosis

The objective of this study was to gain further understanding and elucidation of the fluid dynamic factors and flow-induced mechanisms of the thrombogenic process of platelet deposition onto, and possible subsequent embolization from, the walls of an arterial stenosis. This has been accomplished by measurement of the axial dependence of platelet deposition within a modeled arterial stenosis for a transitional flow and a completely laminar flow field. The stenotic region of the model was collagen-coated to simulate a damaged endothelial lining of an artery. Fluid dynamics within a stenosis was studied using qualitative flow visualization, and was further compared to the in vitro platelet deposition studies. Normalized platelet density (NPD) measurements indicate decreased levels of NPD in the high shear throat region of the stenosis for a Reynolds number of 300 and a drastic increase in NPD at the throat for a Reynolds number of 175. This study provides further understanding of the flow dynamic effects on thrombus development within a stenosis. ^

Design strategies for an artificial neural network based algorithm for automatic incident detection on major arterial streets

Traffic incidents are non-recurring events that can cause a temporary reduction in roadway capacity. They have been recognized as a major contributor to traffic congestion on our nation’s highway systems. To alleviate their impacts on capacity, automatic incident detection (AID) has been applied as an incident management strategy to reduce the total incident duration. AID relies on an algorithm to identify the occurrence of incidents by analyzing real-time traffic data collected from surveillance detectors. Significant research has been performed to develop AID algorithms for incident detection on freeways; however, similar research on major arterial streets remains largely at the initial stage of development and testing. This dissertation research aims to identify design strategies for the deployment of an Artificial Neural Network (ANN) based AID algorithm for major arterial streets. A section of the US-1 corridor in Miami-Dade County, Florida was coded in the CORSIM microscopic simulation model to generate data for both model calibration and validation. To better capture the relationship between the traffic data and the corresponding incident status, Discrete Wavelet Transform (DWT) and data normalization were applied to the simulated data. Multiple ANN models were then developed for different detector configurations, historical data usage, and the selection of traffic flow parameters. To assess the performance of different design alternatives, the model outputs were compared based on both detection rate (DR) and false alarm rate (FAR). The results show that the best models were able to achieve a high DR of between 90% and 95%, a mean time to detect (MTTD) of 55-85 seconds, and a FAR below 4%. The results also show that a detector configuration including only the mid-block and upstream detectors performs almost as well as one that also includes a downstream detector. In addition, DWT was found to be able to improve model performance, and the use of historical data from previous time cycles improved the detection rate. Speed was found to have the most significant impact on the detection rate, while volume was found to contribute the least. The results from this research provide useful insights on the design of AID for arterial street applications.

Monounsaturated fatty acid, carbohydrate intake, and diabetes status are associated with arterial pulse pressure

Background Diabetes is a global epidemic. Cardiovascular disease (CVD) is one of the most prevalent consequences of diabetes. Nutrition is considered a modifiable risk factor for CVD, particularly for individuals with diabetes; albeit, there is little consensus on the role of carbohydrates, proteins and fats for arterial health for persons with or without diabetes. In this study, we examined the association of macronutrients with arterial pulse pressure (APP), a surrogate measure of arterial health by diabetes status and race. Methods Participants were 892 Mexican Americans (MA), 1059 Black, non-Hispanics (BNH) and 2473 White, non-Hispanics (WNH) with and without diabetes of a weighted sample from the National Nutrition and Health Examination Survey (NHANES) 2007-2008. The cross-sectional analysis was performed with IBM-SPSS version 18 with the complex sample analysis module. The two-year sample weight for the sub-sample with laboratory values was applied to reduce bias and approximate a nationally, representative sample. Arterial stiffness was assessed by arterial pulse pressure (APP). Results APP was higher for MA [B = 0.063 (95% CI 0.015 to 0.111), p = 0.013] and BNH [B = 0.044 (95% CI 0.006 to 0.082), p = 0.018] than WNH, controlling for diabetes, age, gender, body mass index (BMI), fiber intake, energy intake (Kcal) and smoking. A two-way interaction of diabetes by carbohydrate intake (grams) was inversely associated with APP [B = -1.18 (95% CI -0.178 to -0.058), p = 0.001], controlling for race, age, gender, BMI, Kcal and smoking. BNH with diabetes who consumed more mono-unsaturated fatty acids (MUFA) than WNH with diabetes had lower APP [B = -0.112 (95%CI-0.179 to -0.045), p = 0.003] adjusting for saturated fatty acids, Kcal, age, gender, BMI and smoking. Conclusion Higher MUFA and carbohydrate intake for persons with diabetes reflecting lower APP may be due to replacement of saturated fats with CHO and MUFA. The associations of APP with diabetes, race and dietary intake need to be confirmed with intervention and prospective studies. Confirmation of these results would suggest that dietary interventions for minorities with diabetes may improve arterial health.

The Effect of Carbohydrate Amount, Quality and Type on Arterial Pulse Pressure in Cuban-Americans with and Without Type 2 Diabetes

Background: Arterial pulse pressure, the difference between systolic and diastolic blood pressure, has been used as an indicator (surrogate measure) of arterial stiffness. High arterial pulse pressure (> 40) has been associated with increased cardiovascular disease and mortality. Several clinical trials have reported that the proportion of calories from carbohydrate has an effect on blood pressure. The primary objective of this study was to assess arterial pulse pressure and its association with carbohydrate quantity and quality (glycemic load) with diabetes status for a Cuban American population. Methods: A single point analysis included 367 participants. There was complete data for 365 (190 with and 175 without type 2 diabetes). The study was conducted in the investigator’s laboratory located in Miami, Florida. Demographic, dietary, anthropometric and laboratory data were collected. Arterial pulse pressure was calculated by the formula systolic minus the diastolic blood pressure. Glycemic load, fructose, sucrose, percent of average daily calories from carbohydrate, fat and protein, grams of fiber and micronutrient intakes were calculated from a validated food frequency questionnaire. Results: The mean arterial pulse pressure was significantly higher in participants with (52.9 ± 12.4) than without (48.6 ± 13.4) type 2 diabetes. The odds of persons with diabetes having high arterial pulse pressure (>40) was 1.85 (95% CI =1.09, 3.13); p=0.023. For persons with type 2 diabetes higher glycemic load was associated with lower arterial pulse pressure. Conclusions: Arterial pulse pressure and diet are modifiable risk factors of cardiovascular disease. Arterial pulse pressure may be associated with carbohydrate intake differently considering diabetes status. Results may be due to individuals with diabetes following dietary recommendations. The findings of this study suggest clinicians take into consideration how medical condition, ethnicity and diet are associated with arterial pulse pressure before developing a medical nutrition therapy plan in collaboration with the client.

Phylloquinone (Vitamin K₁) Intake and Pulse Pressure as a Measure of Arterial Stiffness in Older Adults

This study examined the relationships among ethnicity/race, lifestyle factors, phylloquinone (vitamin K₁) intake, and arterial pulse pressure in a nationally representative sample of older adults from four ethnic/racial groups: non-Hispanic Whites, non-Hispanic Blacks, Mexican Americans, and other Hispanics. This was a cross-sectional study of U.S. representative sample with data from the National Health and Nutrition Examination Surveys, 2007-2008 and 2009-2010 of adults aged 50 years and older (N = 5296). Vitamin K intake was determined by 24-hour recall. Pulse pressure was calculated as the difference between the averages of systolic blood pressure and diastolic blood pressure. Compared to White non-Hispanics, the other ethnic/racial groups were more likely to have inadequate vitamin K₁ intake. Inadequate vitamin K₁ intake was an independent predictor of high arterial pulse pressure. This was the first study that compared vitamin K₁ inadequacy with arterial pulse pressure across ethnicities/races in U.S. older adults. These findings suggest that vitamin K screening may be a beneficial marker for the health of older adults.

Design Strategies for an Artificial Neural Network Based Algorithm for Automatic Incident Detection on Major Arterial Streets

Traffic incidents are non-recurring events that can cause a temporary reduction in roadway capacity. They have been recognized as a major contributor to traffic congestion on our national highway systems. To alleviate their impacts on capacity, automatic incident detection (AID) has been applied as an incident management strategy to reduce the total incident duration. AID relies on an algorithm to identify the occurrence of incidents by analyzing real-time traffic data collected from surveillance detectors. Significant research has been performed to develop AID algorithms for incident detection on freeways; however, similar research on major arterial streets remains largely at the initial stage of development and testing. This dissertation research aims to identify design strategies for the deployment of an Artificial Neural Network (ANN) based AID algorithm for major arterial streets. A section of the US-1 corridor in Miami-Dade County, Florida was coded in the CORSIM microscopic simulation model to generate data for both model calibration and validation. To better capture the relationship between the traffic data and the corresponding incident status, Discrete Wavelet Transform (DWT) and data normalization were applied to the simulated data. Multiple ANN models were then developed for different detector configurations, historical data usage, and the selection of traffic flow parameters. To assess the performance of different design alternatives, the model outputs were compared based on both detection rate (DR) and false alarm rate (FAR). The results show that the best models were able to achieve a high DR of between 90% and 95%, a mean time to detect (MTTD) of 55-85 seconds, and a FAR below 4%. The results also show that a detector configuration including only the mid-block and upstream detectors performs almost as well as one that also includes a downstream detector. In addition, DWT was found to be able to improve model performance, and the use of historical data from previous time cycles improved the detection rate. Speed was found to have the most significant impact on the detection rate, while volume was found to contribute the least. The results from this research provide useful insights on the design of AID for arterial street applications.

Influência das medidas antropométricas sobre a pressão arterial sistêmica de hipertensos cadastrados nas unidades de saúde da família no município de João Pessoa - PB

Este trabalho objetiva avaliar a influência das medidas antropométricas sobre a pressão arterial sistêmica de uma coorte de hipertensos atendidos nas Unidades de Saúde da Família (USF) no município de João Pessoa-PB durante o período de 2008 a 2011. Para verificar as diferenças de níveis pressóricos entre as medidas realizadas nas consultas referentes ao cadastro, e a ultima pressão obtida no período de acompanhamento foram utilizadas informações sobre a identificação, dados clínicos do paciente, fatores de risco e doenças concomitantes. Testes estatísticos de comparabilidade entre grupos foram empregados. Para investigar os padrões de associação dos fatores de risco das variáveis antropométricas e sociodemográficas com a hipertensão dos pacientes, foi empregado o Modelo de Regressão Logística. Foram encontradas associações significativas (p<0,005) da hipertensão com a obesidade e o sedentarismo. Com os resultados desta pesquisa pretende-se: fornecer indicadores sobre a efetividade da Estratégia Saúde da Família; avaliar a qualidade das informações antropométricas; contribuir para a definição de estratégias que garantam o melhoramento dos acompanhamentos dos hipertensos e fornecer elementos científicos de apoio para outras USF e instituições governamentais.

Relação entre desempenho funcional e indicadores de doença arterial obstrutiva periférica em idosas

The presence of peripheral arterial disease (PAD) increases the risk and vulnerability to adverse clinical outcomes in the elderly. Like this, we investigated the relationship between functional performance and indicators of PAD in elderly women. METHODS: Cross-sectional study in which 54 elderly were assessed by questionnaire mini mental state examination, Short Phisical Performance Battery (SPPB), ankle-brachial index (ABI), human activity profile (HAP) and Edinburgh questionnaire. Statistical analysis was performed using ANOVA, t test and Pearson correlation. We considered p <0.05 as significance level. RESULTS: The mean age SPPB and ABI were 69.2 (± 6.9) years, SPPB 9.42 (± 2.55) and ABI 1.04 (± 0.14). The prevalence of PAD was 16.3%. There was a significant correlation between ABI and gait speed (r = 0.75, p = 0.001) and between PAH with SPPB (p = 0.001). CONCLUSIONS: It is suggested that the decline in functional performance in older, expressed in gait velocity component of the SPPB, is related to the presence of PAD

Safety and tolerability considerations in the use of sildenafil for children with pulmonary arterial hypertension.

Sildenafil is a phosphodiesterase type-5 inhibitor approved for treatment of pulmonary arterial hypertension (PAH) in adults. Data from pediatric trials demonstrate a similar acute safety profile to the adult population but have raised concerns regarding the safety of long-term use in children. Interpretation of these trials remains controversial with major regulatory agencies differing in their recommendations - the US Food and Drug Administration recommends against the use of sildenafil for treatment of PAH in children, while the European Medicines Agency supports its use at "low doses". Here, we review the available pediatric data regarding dosing, acute, and long-term safety and efficacy of sildenafil for the treatment of PAH in children.