bluuebunny/crossref_metadata_2025 huggingface dataset:
Grain Sorghum Response to Hybrid, Row Spacing, and Plant Populations along the Upper Texas Gulf Coast
Authors: Fromme Dan D. , Fernandez Carlos J. , Grichar W. James , Jahn Rick L.
Published: July 2012

Grain sorghum (Sorghum bicolorL. Moench) along the upper Texas Gulf Coast is planted at different row spacings, plant populations, and hybrids according to grower preference and location. This study was conducted over a 3-year period (2001 to 2003) to determine the combination of hybrid, row spacing, and plant population on yield and net return per hectare. Below average rainfall for May and June occurred in 2002 and 2003 which resulted in lower yields than 2001. In 2001 and 2003, Dekalb (DK) 54 generally produced higher yields than Asgrow (A) 571 on the 51 cm row spacing. In 2002, no differences in grain sorghum yield were found when comparing both hybrids, plant populations, or row spacing with the exception of A 571 planted at 148,000 plants/ha on 102 cm centers which resulted in a yield reduction of at least 25%. When net returns were compared regardless of year or rainfall received, net dollar value per hectare tended to be higher for the 51 cm row spacing, especially with A 571.
A Theoretical Approach to Predict the Fatigue Life of Flexible Pipes
Authors: de Sousa José Renato M. , de Sousa Fernando J. M. , de Siqueira Marcos Q. , Sagrilo Luís V. S. , de Lemos Carlos Alberto D.
Published: July 2012

This paper focuses on a theoretical approach to access the fatigue life of flexible pipes. This methodology employs functions that convert forces and moments obtained in time‐domain global analyses into stresses in their tensile armors. The stresses are then processed by well‐known cycle counting methods, and S-N curves are used to evaluate the fatigue damage at several points in the pipe’s cross‐section. Finally, Palmgren‐Miner linear damage hypothesis is assumed in order to calculate the accumulated fatigue damage. A study on the fatigue life of a flexible pipe employing this methodology is presented. The main points addressed in the study are the influence of friction between layers, the effect of the annulus conditions, the importance of evaluating the fatigue life in various points of the pipe’s cross‐section, and the effect of mean stresses. The results obtained suggest that the friction between layers and the annulus conditions strongly influences the fatigue life of flexible pipes. Moreover, mean stress effects are also significant, and at least half of the wires in each analyzed section of the pipe must be considered in a typical fatigue analysis.
Antibacterial Activity of Daucus crinitus Essential Oils along the Vegetative Life of the Plant
Authors: Bendiabdellah Amel , Dib Mohammed El Amine , Meliani Nawel , Muselli Alain , Nassim Djabou , Tabti Boufeldja , Costa Jean
Published: July 2012

The essential oils from the aerial parts of Daucus crinitus Desf. were analyzed at three developmental stages (early vegetative, early flowering, and full flowering). Oil yield was found to vary depending on the stage of development, and the highest content of oil (0.15% w/w) was obtained at full flowering. The chemical composition of essential oils studied by GC and GC‐MS showed a total of 71 compounds: 27 aliphatic compounds, 18 sesquiterpene hydrocarbons, 9 hydrocarbons monoterpene, 5 oxygenated monoterpenes, 5 phenolic compounds, 4 oxygenated sesquiterpenes, 2 oxygenated diterpenes, and 01 diterpene hydrocarbons. Whatever the analyzed stage, phenolic compounds were the most abundant group. Their level significantly increased during ripening and varied from 36.4 to 82.1%. Antimicrobial activities of oils were tested on four different microorganisms. The oils of various phenological stages showed high activity against Candida albicans (30 mm) and Staphylococcus aureus (11–28 mm) bacteria strains which are deemed very dangerous and very difficult to eliminate. Thus, they represent an inexpensive source of natural antibacterial substances that may potentially be used in pathogenic systems.
Dielectric and Excess Dielectric Constants of Acetonitrile + Butyl Amine, + Ethylamine, and + Methylamine at 303, 313, and 323 K
Authors: Ramana Ch. V. V. , Kumar A. B. V. Kiran , Kumar M. Ashok , Moodley M. K.
Published: July 2012

The dielectric constants and excess dielectric constants of the binary systems: acetonitrile + butyl amine, + ethylamine and + methylamine have been studied at 303, 313, and 323 K temperatures and over the complete mole fraction range. The dielectric constants for these mixtures were measured using a microcontroller based system. The results are positive over the entire range of composition. Symmetrical curves were observed for the systems in which the maximum occurs approximately at 0.7‐mole fraction of acetonitrile. The results are discussed in terms of intermolecular interactions. The investigation of dielectric constant of mixed solvents bearing amines aims at better comprehension of their biological, chemical, pharmaceutical, technological, and laboratory applications.
Distribution and Risk Assessment of Polycyclic Aromatic Hydrocarbons in Vegetables Grown in Pakistan
Authors: Ashraf Mohammad W. , Taqvi Syed Iqleem H. , Solangi Amber R. , Qureshi Umair A.
Published: July 2012

Distribution and risk assessment of eight priority polycyclic aromatic hydrocarbons (PAHs) contents have been examined in different varieties of vegetables grown in Pakistan. The results showed that the total PAH contents were higher for root vegetables like potato and carrot (~13 μg/kg) and relatively lower for turnip
(10.9 μg/kg), respectively while for the fruit vegetables, all the peels were found to be more contaminated than cores. The ratio of total PAH concentrations in peels with respect to those of cores is found to be 1.45, 1.26, 1.31, 1.44, 1.40, and 1.36 for potato, turnip, carrot, eggplant, cucumber, and bitter gourd, respectively. For leafy vegetables, cabbage showed maximum PAH (11.6 μg/kg) as compared to the cores of fruit vegetables. Among individual PAH congeners, anthracene showed higher levels in all vegetables. For benzo(a)anthracene, maximum concentration (3.44 ± 2.10 μg/kg) was encountered in turnip cores. Highest benzo(e)pyrene concentration was found in potato (3.19 ± 1.67 μg/kg) followed by turnip (2.74 ± 1.22 μg/kg). Benzo(b)fluoranthene and benzo(k)fluoranthene showed relatively lower levels in all samples studied. All the concentrations of PAHs are presented on a fresh weight basis. The results of human exposure of PAH by consumption of these vegetables showed that cumulative dietary exposure of Pakistani population to PAHs from vegetables ranges from 0.25 μg/p/d to 1.16 μg/p/d.
Impaired IFN-α Production by Plasmacytoid Dendritic Cells Favors Regulatory T-cell Expansion That May Contribute to Breast Cancer Progression
Authors: Sisirak Vanja , Faget Julien , Gobert Michael , Goutagny Nadège , Vey Nelly , Treilleux Isabelle , Renaudineau Sarah , Poyet Gaelle , Labidi-Galy Sana Intidhar , Goddard-Leon Sophie , Durand Isabelle , Le Mercier Isabelle , Bajard Agathe , Bachelot Thomas , Puisieux Alain , Puisieux Isabelle , Blay Jean-Yves , Ménétrier-Caux Christine , Caux Christophe , Bendriss-Vermare Nathalie
Published: July 2012

Infiltration and dysfunction of immune cells have been documented in many types of cancers. We previously reported that plasmacytoid dendritic cells (pDC) within primary breast tumors correlate with an unfavorable prognosis for patients. The role of pDC in cancer remains unclear but they have been shown to mediate immune tolerance in other pathophysiologic contexts. We postulated that pDC may interfere with antitumor immune response and favor tolerance in breast cancer. The present study was designed to decipher the mechanistic basis for the deleterious impact of pDC on the clinical outcome. Using fresh human breast tumor biopsies (N = 60 patients), we observed through multiparametric flow cytometry increased tumor-associated (TA) pDC (TApDC) rates in aggressive breast tumors, i.e., those with high mitotic index and the so-called triple-negative breast tumors (TNBT). Furthermore, TApDC expressed a partially activated phenotype and produced very low amounts of IFN-α following toll-like receptor activation in vitro compared with patients' blood pDC. Within breast tumors, TApDC colocalized and strongly correlated with TA regulatory T cells (TATreg), especially in TNBT. Of most importance, the selective suppression of IFN-α production endowed TApDC with the unique capacity to sustain FoxP3+ Treg expansion, a capacity that was reverted by the addition of exogenous IFN-α. These findings indicate that IFN-α–deficient TApDC accumulating in aggressive tumors are involved in the expansion of TATreg in vivo, contributing to tumor immune tolerance and poor clinical outcome. Thus, targeting pDC to restore their IFN-α production may represent an attractive therapeutic strategy to overcome immune tolerance in breast cancer. Cancer Res; 72(20); 5188–97. ©2012 AACR.
Platelets and P-Selectin Control Tumor Cell Metastasis in an Organ-Specific Manner and Independently of NK Cells
Authors: Coupland Lucy A. , Chong Beng H. , Parish Christopher R.
Published: July 2012

The prometastatic role of platelets has long been recognized with proposed mechanisms of action including shielding tumor cells from natural killer (NK) cell destruction and aiding endothelial attachment and extravasation of tumor cells with platelet P-selectin being implicated in these processes. However, many aspects of the prometastatic function of platelets remain unclear. In this study, we used mouse models of metastatic breast cancer and melanoma to investigate the platelet effect, focusing on organ specificity, the relationship with NK cells and the relative importance of platelet-derived versus endothelial-derived P-selectin. We found that platelets promote lung metastasis in the absence of NK cells in both acute and spontaneous metastasis models. In addition, the prometastatic action of platelets was found to be organ specific, clearly enhancing lung metastasis but not affecting B16F1 liver metastasis, in fact, liver metastasis was enhanced in the absence of platelets. Furthermore, the profound antimetastatic activity of NK cells was equally effective in the presence or absence of platelets and chronologically distinct from the prometastatic role of platelets. Finally, it was shown that endothelial-derived P-selectin is just as important as platelet-derived P-selectin in promoting lung metastasis and also plays an important role in liver metastasis. Taken together, our findings help clarify the roles of platelets, NK cells and P-selectin in metastasis, and they identify P-selectin as an attractive therapeutic target for preventing metastasis in multiple organs. Cancer Res; 72(18); 4662–71. ©2012 AACR.
Systemic Combination Virotherapy for Melanoma with Tumor Antigen-Expressing Vesicular Stomatitis Virus and Adoptive T-cell Transfer
Authors: Rommelfanger Diana M. , Wongthida Phonphimon , Diaz Rosa M. , Kaluza Karen M. , Thompson Jill M. , Kottke Timothy J. , Vile Richard G.
Published: July 2012

Oncolytic virotherapy offers the potential to treat tumors both as a single agent and in combination with traditional modalities such as chemotherapy and radiotherapy. Here we describe an effective, fully systemic treatment regimen, which combines virotherapy, acting essentially as an adjuvant immunotherapy, with adoptive cell transfer (ACT). The combination of ACT with systemic administration of a vesicular stomatitis virus (VSV) engineered to express the endogenous melanocyte antigen glycoprotein 100 (gp100) resulted in regression of established melanomas and generation of antitumor immunity. Tumor response was associated with in vivo T-cell persistence and activation as well as treatment-related vitiligo. However, in a proportion of treated mice, initial tumor regressions were followed by recurrences. Therapy was further enhanced by targeting an additional tumor antigen with the VSV-antigen + ACT combination strategy, leading to sustained response in 100% of mice. Together, our findings suggest that systemic virotherapy combined with antigen-expressing VSV could be used to support and enhance clinical immunotherapy protocols with adoptive T-cell transfer, which are already used in the clinic. Cancer Res; 72(18); 4753–64. ©2012 AACR.
MicroRNA-21 Modulates the Levels of Reactive Oxygen Species by Targeting SOD3 and TNFα
Authors: Zhang Xiangming , Ng Wooi-Loon , Wang Ping , Tian LinLin , Werner Erica , Wang Huichen , Doetsch Paul , Wang Ya
Published: July 2012

MicroRNA-21 (miR-21) is an oncomir overexpressed in most human tumors in that it promotes malignant growth and progression by acting on multiple targets. Here, we broaden the impact of miR-21 in cancer by showing that it regulates the formation of reactive oxygen species (ROS) that promote tumorigenesis. Key targets of miR-21 in mediating this function were SOD3 and TNFα. We found that miR-21 inhibited the metabolism of superoxide to hydrogen peroxide, produced either by endogenous basal activities or exposure to ionizing radiation (IR), by directing attenuating SOD3 or by an indirect mechanism that limited TNFa production, thereby reducing SOD2 levels. Importantly, both effects contributed to an elevation of IR-induced cell transformation. Our findings, therefore, establish that miR-21 promotes tumorigenesis to a large extent through its regulation of cellular ROS levels. Cancer Res; 72(18); 4707–13. ©2012 AACR.
Clinical Trial: Transcutaneous Interferential Electrical Stimulation in Individuals with Irritable Bowel Syndrome – A Prospective Double-Blind Randomized Study
Authors: Çoban Şahin , Akbal Erdem , Köklü Seyfettin , Köklü Gülşah , Ulaşlı Murat Alper , Erkeç Serap , Aktaş Bora , Yüksel Osman , Koçak Erdem , Erdem Hatice Rana
Published: July 2012

Background: The exact etiology of irritable bowel syndrome (IBS) remains unclear. Curative treatment is not available and current treatment modalities are mainly directed against the predominant symptoms. There are a few studies reporting the beneficial effects of transcutaneous electrical stimulation in patients with chronic constipation, gastroparesis, and functional dyspepsia. Aim: To investigate whether transcutaneous electrical stimulation is an effective procedure in IBS patients. Methods: IBS patients were randomly placed in vacuum interferential current (IFC) and placebo groups. Both treatments consisted of 12 sessions administered over 4 weeks. Symptoms due to IBS were documented via questionnaires, including the IBS Global Assessment of Improvement Scale, numeric rating scales, visual analogue scale, and IBS Quality of Life Scale at the beginning of, end of, and 1 month after the treatment. Results: Patients in the therapy (29 cases) and placebo (29 cases) groups were homogeneous with respect to demographic data and gastrointestinal system symptoms. When compared to the beginning scores, severity of abdominal discomfort, bloating, and abdominal distension and rumbling improved significantly in either interference or placebo groups at both the end of treatment and 1 month after treatment. In the IFC group, severity of symptoms continued to decrease significantly at 1 month after treatment when compared to scores at just the end of treatment, whereas in the placebo group severity of these symptoms did not change significantly on numeric severity scales. Also, the visual analogue scale of the first month after treatment continued to decrease significantly when compared to the level at the end of treatment in the IFC group. Total quality score increased significantly in the IFC group. Conclusions: Vacuum IFC therapy can significantly improve symptoms and quality of life in patients with IBS. It may represent a novel treatment modality for drug-refractory IBS patients.
A Competing Risks Model in Early Screening for Preeclampsia
Authors: Wright David , Akolekar Ranjit , Syngelaki Argyro , Poon Leona C.Y. , Nicolaides Kypros H.
Published: July 2012

Objective: It was the aim of this study to develop models for the prediction of preeclampsia (PE) based on maternal characteristics and biophysical markers at 11–13 weeks’ gestation in which gestation at the time of delivery for PE is treated as a continuous variable. Methods: This was a screening study of singleton pregnancies at 11–13 weeks including 1,426 (2.4%) cases that subsequently developed PE and 57,458 cases that were unaffected by PE. We developed a survival time model for the time of delivery for PE in which Bayes’ theorem was used to combine the prior information from maternal characteristics with the uterine artery pulsatility index (PI) and the mean arterial pressure (MAP), using multiple of the median values. Results: The risk for PE increased with maternal age, weight, Afro-Caribbean and South Asian racial origin, previous pregnancy with PE, conception by in vitro fertilization and a medical history of chronic hypertension, type 2 diabetes mellitus as well as systemic lupus erythematosus or antiphospholipid syndrome. In pregnancies with PE, there was an inverse correlation between multiple of the median values of the uterine artery PI and MAP with gestational age at delivery. Screening by maternal characteristics, uterine artery PI and MAP detected 90% of PE cases requiring delivery before 34 weeks and 57% of all PE cases at a fixed false-positive rate of 10%. Conclusions: A new model has been developed for effective first-trimester screening for PE.
Birthweight with Gestation and Maternal Characteristics in Live Births and Stillbirths
Authors: Poon Leona C.Y. , Volpe Nicola , Muto Brunella , Syngelaki Argyro , Nicolaides Kypros H.
Published: July 2012

Objective: To establish a normal range of birthweight with gestational age (GA) at delivery and examine the contribution of maternal characteristics in defining growth restriction in stillbirths. Methods: In 69,895 normal singleton pregnancies, regression analysis was used to determine the association of birthweight with GA and maternal characteristics. The proportion of 290 stillbirths classified as small for GA depending on inclusion or exclusion of maternal characteristics was determined. Results: In normal pregnancies, there was a polynomial association between birthweight and GA. Birthweight increased with maternal weight, height and parity and was lower in Africans and South Asians than in Caucasians. Birthweight for GA was reduced in antepartum stillbirths (n = 243; p < 0.0001) but not in intrapartum stillbirths (n = 47; p = 0.334). There was no significant difference in the proportion of antepartum stillbirths with birthweight below the 10th percentile when birthweight was corrected for GA only compared to correction for GA and maternal characteristics (53.1 vs. 54.3%). The birthweight was below the 10th percentile in 71.8% of antepartum stillbirths at <32 weeks’ gestation, in 47.2% at 33–36 weeks and in 31.5% at ≥37 weeks. Conclusion: Correction of birthweight for maternal characteristics does not alter the proportion of stillbirths that are small for GA.
Stress-Induced Cardiomyopathy in Sweden: Evidence for Different Ethnic Predisposition and Altered Cardio-Circulatory Status
Authors: Schultz Tomas , Shao Yangzhen , Redfors Björn , Bergmann Sverrisdóttir Yrsa , Råmunddal Truls , Albertsson Per , Matejka Göran , Omerovic Elmir
Published: July 2012

Background: In this paper, we report about new insights regarding clinical course, long-term outcome, ethnic/genetic predisposition and cardio-circulatory status in the large stress-induced cardiomyopathy (SIC) cohort from Sweden. Methods and Results: We have included 115 consecutive SIC patients between January 2005 and January 2010 at Sahlgrenska University Hospital in Gothenburg. Hemodynamic status and sympathetic nerve activity were evaluated and compared with those of healthy controls. Mean age was 64, and 14% were males. Thirty-day and 3-year mortality was 6 and 10%, respectively. Eleven percent had ischemic heart disease, 3% developed thromboembolic complications, 6% had cardiac arrest and 14% developed cardiogenic shock. The great majority of SIC patients (93%) were ethnic Swedes. In three families, several close relatives developed SIC. Fourteen percent developed two or more episodes of SIC. Hemodynamic evaluation has shown subnormal systemic vascular resistance, 22% lower sympathetic activity and preserved cardiac output in SIC patients. Conclusions: SIC affects both men and women of different ages and is associated with significant short- and long-term mortality. There is a strong signal for the presence of ethnic/genetic predisposition to develop SIC. Sympathetic activity and systemic vascular resistance are lower in SIC patients, suggesting that SIC is a cardio-circulatory phenomenon.
Cardiac Rehabilitation to Improve Physical Functioning in Refractory Angina: A Pilot Study
Authors: Asbury Elizabeth A. , Webb Carolyn M. , Probert Heather , Wright Christine , Barbir Mahmoud , Fox Kim , Collins Peter
Published: July 2012

Objectives: Refractory angina patients suffer debilitating chest pain despite optimal medical therapy and previous cardiovascular intervention. Cardiac rehabilitation is often not prescribed due to a lack of evidence regarding potential efficacy and patient suitability. A randomised controlled study was undertaken to explore the impact of cardiac rehabilitation on cardiovascular risk factors, physical ability, quality of life and psychological morbidity among refractory angina sufferers. Methods: Forty-two refractory angina patients (65.1 ± 7.3 years) were randomly assigned to an 8-week Phase III cardiac rehabilitation program or symptom diary control. Physical assessment, Progressive Shuttle Walk test, Hospital Anxiety and Depression Scale, Health Anxiety Questionnaire, the York Angina Beliefs scale, ENRICHD Social Support Instrument and SF-36 were completed before and after intervention and at 8-week follow-up. Results: Following cardiac rehabilitation, patients demonstrated improved physical ability compared with controls in Progressive Shuttle Walk level attainment (p = 0.005) and total distance covered (p = 0.015). Angina frequency and severity remained unchanged in both groups, with the control demonstrating worsening SF-36 pain scale (63.43 ± 22.28 vs. 55.46 ± 23.98, p = 0.025). Cardiac rehabilitation participants showed improved Health Anxiety Questionnaire reassurance (1.71 ± 1.72 vs. 1.14 ± 1.23, p = 0.026) and York Beliefs anginal threat perception (12.42 ± 4.58 vs. 14.35 ± 4.73, p = 0.05) after cardiac rehabilitation. Physical measures were broadly unaffected. Conclusions: Cardiac rehabilitation can be prescribed to improve physical ability without affecting angina frequency or severity among patients with refractory angina.
Liver Transplantation in Nonalcoholic Steatohepatitis Is Associated with High Mortality and Post-Transplant Complications: A Single-Center Experience
Authors: Heuer Matthias , Kaiser Gernot M. , Kahraman Alisan , Banysch Mark , Saner Fuat H. , Mathé Zoltan , Gerken Guido , Paul Andreas , Canbay Ali , Treckmann Jürgen W.
Published: July 2012

Background/Aims: Non-alcoholic fatty liver disease (NAFLD) with its progressive form nonalcoholic steatohepatitis (NASH) is the most common chronic liver disease in western countries which is associated with end-stage liver disease and hepatocellular carcinoma (HCC). This entity is a consistently increasing indication for transplantation. However, data about postsurgery outcome and complications are still limited. Patients and Methods: Records of 432 consecutive transplanted patients between October 2007 and January 2011 were investigated retrospectively. Forty transplants were performed due to NASH-induced cirrhosis. Perioperative courses and short- and long-term outcomes were analyzed. Results: The NAFLD population consisted of 16 women and 24 men with a mean age of 55 years. The median MELD score was 27 at the time of liver transplantion. BMI before surgery ranged from 21 to 45 (mean 31). Sixteen of the initial 40 patients are still alive. Patients with sustained obesity and features of the metabolic syndrome had a worse 1-year mortality rate of 42%. Conclusions: A significant number of liver transplantations in our center was performed due to NASH; transplantation in this cohort was associated with high mortality and postoperative complications, most likely due to associated obesity and diabetes. Weight reduction prior to surgery may lead to a better outcome.
Orexin A Expression and Promoter Methylation in Patients with Cannabis Dependence in Comparison to Nicotine-Dependent Cigarette Smokers and Nonsmokers
Authors: Rotter Andrea , Bayerlein Kristina , Hansbauer Max , Weiland Judith , Sperling Wolfgang , Kornhuber Johannes , Biermann Teresa
Published: July 2012

Background: The orexins (hypocretins) are neuropeptides with an origin in the lateral hypothalamus. They have been found to be crucial within the context of drug craving, withdrawal und relapse. Methods: Therefore, orexin A gene expression and promoter methylation in peripheral blood cells of 77 subjects [36 with tetrahydrocannabinol (THC) dependence, 20 nicotine-dependent cigarette smokers and 21 nonsmokers] were assessed by quantitative real-time PCR and methylation-specific digestion PCR. Results: There was a statistically significant difference in orexin A expression between the three groups [p = 0.000, F = 131.4, d.f. = 2, analysis of variance (ANOVA)]. Orexin A gene expression was statistically significantly correlated with the Satisfaction with Life Scale (r = –0.28, p = 0.018), a visual analogue scale of craving (r = 0.734, p = 0.000) and three subscales of the World Health Organization Alcohol, Smoking and Substance Involvement Screening Test, i.e. nicotine consumption (r = 0.388, p = 0.001), alcohol consumption (r = 0.354, p = 0.002) and cannabis consumption (r = 0.783, p = 0.000). The mean promoter methylation (as a percentage) was not statistically related to orexin gene expression. However, there was a statistically significant difference in promoter methylation with regard to body mass index in general (F = 2.37, d.f. = 54, p = 0.016, ANOVA). Conclusions: Orexin might be a possible target in THC as well as nicotine dependence, taking into account the effect of THC on energy homeostasis in the circuit of reward and motivation and its impact on appetite and body weight.
Relationship of Hemoglobin and Hematocrit to Systolic Function in Advanced Heart Failure
Authors: Guglin Maya , Darbinyan Nellie
Published: July 2012

Objectives: The dataset from the Evaluation Study of Congestive Heart Failure and Pulmonary Artery Catheterization Effectiveness (ESCAPE) trial provides a rare opportunity to evaluate the whole spectrum of associations of hemoglobin (HB) and hematocrit (HCT) in heart failure (HF). In that trial, subjective and objective data were recorded at multiple time points when HB and HCT were also measured. We investigated the relationship between anemia and ventricular systolic function. Methods: A limited access dataset from the ESCAPE trial, provided by the National Heart, Lung and Blood Institute, was analyzed. Linear regression analysis, correlation coefficients and Student’s t test were utilized. Results: Besides the known association of anemia with poor prognosis, more severe symptoms, decreased functional capacity and impaired kidney function, we found a significant and very consistent inverse correlation between HB and HCT and ventricular contractility. Both left ventricular ejection fraction and right ventricular fractional area change improved with a decrease in HB and vice versa. We hypothesize that this effect can result from a change in viscosity, which decreases with a decrease in HCT, and may facilitate adaptation of the heart to a volume overload state accompanied by hemodilution. Conclusions: In HF, anemia is associated with poor prognosis and functional impairment, but also with mildly improved systolic function. It may represent an adaptive reaction to congestion.
Physiologic Consequences of Glucose Transport and Phosphoenolpyruvate Node Modifications in Bacillus subtilis 168
Authors: Cabrera-Valladares Natividad , Martínez Luz M. , Flores Noemí , Hernández-Chávez Georgina , Martínez Alfredo , Bolívar Francisco , Gosset Guillermo
Published: July 2012

The phosphoenolpyruvate (PEP) node is an important carbon distribution point in the central metabolic networks; therefore, its modification is a common strategy employed for developing microbial production strains. In this study, mutants of Bacillus subtilis 168 were generated with deletions of pykA (which encodes pyruvate kinase), ptsG (which encodes the glucose-specific IICBAGlc component) or the ptsGHI operon [which encodes IICBAGlc, HPr protein and enzyme I from the PEP:sugar phosphotransferase system (PTS)]. These modifications caused a reduction in the initial rate of [14C]-glucose import, corresponding to 10.99, 2.83 and 0.50% of that found in B. subtilis 168 for strains with inactive pykA, ptsG or ptsGHI genes, respectively. Characterization of derivative strains lacking 3-dehydroquinate synthase activity showed that inactivation of pykA leads to an 8-fold increase in carbon flow to the common aromatic pathway. Quantitative real-time PCR analyses of 76 genes from several functional classes revealed a carbon starvation transcriptional pattern that includes a partial gluconeogenic response and overexpression of genes encoding non-PTS glucose importers in the strains lacking functional pykA, ptsG or ptsGHI genes. A transcriptional response consistent with pyruvate limitation was also detected, which includes upregulation of genes encoding malic enzymes that generate pyruvate from malate.
Biological variation in tPA-induced plasma clot lysis time
Authors: Talens Simone , Malfliet Joyce J. M. C. , Rudež Goran , Spronk Henri M. H. , Janssen Nicole A. H. , Meijer Piet , Kluft Cornelis , de Maat Moniek P. M. , Rijken Dingeman C.
Published: July 2012

Hypofibrinolysis is a risk factor for venous and arterial thrombosis, and can be assessed by using a turbidimetric tPA-induced clot lysis time (CLT) assay. Biological variation in clot lysis time may affect the interpretation and usefulness of CLT as a risk factor for thrombosis. Sufficient information about assay variation and biological variation in CLT is not yet available. Thus, this study aimed to determine the analytical, within-subject and between-subject variation in CLT. We collected blood samples from 40 healthy individuals throughout a period of one year (average 11.8 visits) and determined the CLT of each plasma sample in duplicate. The mean (± SD) CLT was 83.8 (± 11.1) minutes. The coefficients of variation for total variation, analytical variation, within-subject variation and between-subject variation were 13.4%, 2.6%, 8.2% and 10.2%, respectively. One measurement can estimate the CLT that does not deviate more than 20% from its true value. The contribution of analytical variation to the within-subject variation was 5.0%, the index of individuality was 0.84 and the reference change value was 23.8%. The CLT was longer in the morning compared to the afternoon and was slightly longer in older individuals (> 40 years) compared to younger (≤40 years) individuals. There was no seasonal variation in CLT and no association with air pollution. CLT correlated weakly with fibrinogen, C-reactive protein, prothrombin time and thrombin generation. This study provides insight into the biological variation of CLT, which can be used in future studies testing CLT as a potential risk factor for thrombosis.
Alteration of the platelet transcriptome in chronic kidney disease
Authors: Plé Hélène , Maltais Manon , Corduan Aurélie , Rousseau Guy , Madore François , Provost Patrick
Published: July 2012

Bleeding and thrombotic disorders are major complications affecting patients with chronic kidney disease (CKD). Exposure of circulating platelets to uraemic toxins and contact with artificial surfaces during dialysis induce platelet abnormalities and alter the platelet proteome. We hypothesised that these changes may be subsequent to changes in the composition and/or regulation of the platelet transcriptome. In this study, we investigated the circulating platelets of 10 CKD patients (i.e. five chronic haemodialysis patients and five stage 4 CKD uraemic patients) and five age- and sex-matched healthy subjects. We observed an alteration of the platelet messenger RNA (mRNA) and microRNA transcriptome in CKD patients. Impaired in uraemic platelets, the levels of some mRNAs and of most microRNAs appeared to be corrected by dialysis, which is consistent with a beneficial effect of dialysis and a mRNA regulatory role of platelet microRNAs. Reduced in platelets of uraemic patients, phosphatidylcholine transfer protein (PCTP) and WD repeat-containing protein 1 (WDR1) were found to be regulated by microRNAs, the latter of which involving hsa-miR-19b, a microRNA increased in platelets of uraemic patients and involved in platelet reactivity. These results suggest that an alteration of microRNA-based mRNA regulatory mechanisms may underlie the platelet response to uremia and entail the development of platelet-related complications in CKD.
Viewpoint: Central adjudication of myocardial infarction in outcome-driven clinical trials – Common patterns in TRITON, RECORD, and PLATO?
Authors: Atar Dan , Serebruany Victor L.
Published: July 2012

Central adjudication in randomised controlled outcome-driven trials represents a traditional approach to maintain data integrity by applying uniformed rules for assessment of clinical events. It was the purpose of this investigation to determine the patterns of myocardial infarction (MI) adjudication in the TRITON, RECORD, and PLATO trials. We were matching centrally-adjudicated MI’s (CAMI’s) from the official trial publication with the site-reported MI (SRMI’s) count from the Food and Drug Administration’s secondary analyses for the investigational compounds prasugrel (TRITON), rosiglitazone (RECORD), and ticagrelor (PLATO). CAMI numbers showed a remarkable discrepancy to SRMI’s by more than a doubling of the difference: from 72 to 145 events in TRITON favoring prasugrel (from a hazard ratio [HR]=0.76, p=0.08; to a HR=0.76, p<0.001), and from 44 to 89 events in favour of ticagrelor in PLATO (from a HR=0.94, p=0.095; to a HR=0.84, p<0.001). In contrast, in the RECORD trial, the CAMI count was less than the SRMI count (from 24 to 8 events, from a HR=1.42, p=0.93; to a HR=1.14, p=0.96), in this case diminishing cardiovascular hazards in favour of rosiglitazone. In conclusion, central adjudication in the TRITON, the RECORD, and the PLATO trial turned out to have a critical impact on study outcomes. Trial publications should in the future include site-reported major efficacy and safety endpoints to preserve data integrity. The regulatory authorities should consider independent audits when there is a major disagreement between centrally adjudicated and site reported events influencing the results of a major clinical trial.
Fibrinogen gene regulation
Authors: Fish Richard J. , Neerman-Arbez Marguerite
Published: July 2012

The Aα, Bβ and γ polypeptide chains of fibrinogen are encoded by a three gene cluster on human chromosome four. The fibrinogen genes (FGB-FGA-FGG) are expressed almost exclusively in hepatocytes where their output is coordinated to ensure a sufficient mRNA pool for each chain and maintain an abundant plasma fibrinogen protein level. Fibrinogen gene expression is controlled by the activity of proximal promoters which contain binding sites for hepatocyte transcription factors, including proteins which influence fibrinogen transcription in response to acute-phase inflammatory stimuli. The fibrinogen gene cluster also contains cis regulatory elements; enhancer sequences with liver activities identified by sequence conservation and functional genomics. While the transcriptional control of this gene cluster is fascinating biology, the medical impetus to understand fibrinogen gene regulation stems from the association of cardiovascular disease risk with high level circulating fibrinogen. In the general population this level varies from about 1.5 to 3.5 g/l. This variation between individuals is influenced by genotype, suggesting there are genetic variants contributing to fibrinogen levels which reside in fibrinogen regulatory loci. A complete picture of how fibrinogen genes are regulated will therefore point towards novel sources of regulatory variants. In this review we discuss regulation of the fibrinogen genes from proximal promoters and enhancers, the influence of acute-phase stimulation, post-transcriptional regulation by miRNAs and functional regulatory variants identified in genetic studies. Finally, we discuss the fibrinogen locus in light of recent advances in understanding chromosomal architecture and suggest future directions for researching the mechanisms that control fibrinogen expression.
Dysregulated coagulation associated with hypofibrinogenaemia and plasma hypercoagulability: Implications for identifying coagulopathic mechanisms in humans
Authors: Marchi Rita , Walton Bethany L. , McGary Colleen S. , Lin Feng-Chang , Ma Alice D. , Pawlinski Rafal , Mackman Nigel , Campbell Robert A. , Di Paola Jorge , Wolberg Alisa S.
Published: July 2012

Identifying coagulation abnormalities in patients with combined bleeding and thrombosis history is clinically challenging. Our goal was to probe the complexity of dysregulated coagulation in humans by characterizing pathophysiologic mechanisms in a patient with both bleeding and thrombosis. The patient is a 56-year-old female with a history of haematomas, poor wound healing, and thrombosis (retinal artery occlusion and transient cerebral ischaemia). She had a normal activated partial thromboplastin time, prolonged thrombin and reptilase times, and decreased functional and antigenic fibrinogen levels, and was initially diagnosed with hypodysfibrinogenaemia. This diagnosis was supported by DNA analysis revealing a novel FGB mutation (c.656A>G) predicting a Q189R mutation in the mature chain that was present in the heterozygote state. However, turbidity analysis showed that purified fibrinogen polymerisation and degradation were indistinguishable from normal, and Bβ chain subpopulations appeared normal by two-dimensional difference in-gel electrophoresis, indicating the mutated chain was not secreted. Interestingly, plasma thrombin generation testing revealed the patient’s thrombin generation was higher than normal and could be attributed to elevated levels of factor VIII (FVIII, 163–225%). Accordingly, in an arterial injury model, hypofibrinogenaemic mice (Fgn+/−) infused with factor VIII demonstrated significantly shorter vessel occlusion times than saline-infused Fgn+/− mice. Together, these data associate the complex bleeding and thrombotic presentation with combined hypofibrinogenaemia plus plasma hypercoagulability. These findings suggest previous cases in which fibrinogen abnormalities have been associated with thrombosis may also be complicated by co-existing plasma hypercoagulability and illustrate the importance of “global” coagulation testing in patients with compound presentations.
Ferritin Is Independently Associated With the Presence of Coronary Artery Calcium in 12 033 Men
Authors: Sung Ki-Chul , Kang Seok-Min , Cho Eun-Joo , Park Jeong Bae , Wild Sarah H. , Byrne Christopher D.
Published: July 2012

Objective—

Ferritin concentrations are often increased in patients with metabolic syndrome and type 2 diabetes mellitus, but few reports have examined the associations between ferritin and atherosclerosis. We investigated whether any relationship between ferritin and coronary artery calcium score (CACS) >0 (as a marker of atherosclerosis) was independent of potential confounders, such as iron-binding capacity (transferrin), low-grade inflammation, and cardiovascular risk factors.

Methods and Results—

Data were analyzed from a South Korean occupational cohort of 12 033 men who underwent a cardiac computed tomography estimation of CACS and measurements of multiple cardiovascular risk factors. One-thousand three- hundred-fifteen of 12 033 (11.2%) subjects had a CACS >0. For people with a CACS >0, median (interquartile range) ferritin concentration was 196.8 (136.3–291.9) compared with 182.2 (128.1–253.6) in people with a CACS=0;
P
<0.001. In the highest ferritin quartile, 14.7% (442/3008) of subjects had a CACS >0 compared with 9.7% (292/3010) in the lowest quartile (
P
<0.0001). With increasing ferritin quartiles, there were also higher proportions of people with diabetes mellitus (
P
<0.0001), hypertension (
P
<0.0001), coronary heart disease (
P
=0.003), and a Framingham Risk Score >10% (
P
<0.0001). In logistic regression modeling with CACS >0 as the outcome, ferritin but not transferrin was independently associated with CACS >0 (odds ratio for highest quartile versus lowest quartile, 1.66 [95% CI, 1.3–1.98];
P
=0.0001).

Conclusion—

Increased ferritin concentrations are associated with the presence of a marker of early coronary artery atherosclerosis, independently of traditional cardiovascular risk factors including Framingham risk score, transferrin, preexisting vascular disease, diabetes mellitus, metabolic syndrome factors, and low-grade inflammation.
Immunization With a Combination of 2 Peptides Derived From the C5a Receptor Significantly Reduces Early Atherosclerotic Lesion in
Ldlr

tm1Her

Apob

tm2Sgy

J Mice
Authors: Lu Xinjie , Xia Min , Endresz Valeria , Faludi Ildiko , Mundkur Lakshmi , Gonczol Eva , Chen Daxin , Kakkar Vijay V.
Published: July 2012

Objective—

The goal of this study was to assess whether immunization of
Ldlr

tm1Her

Apob

tm2Sgy

J mice with 2 peptides located at the N-terminus of the C5a receptor (C5aR), either alone or in combination, is effective in reducing atherosclerotic lesions.

Methods and Results—

Five- to 6-week-old female
Ldlr

tm1Her

Apob

tm2Sgy

J mice were immunized using a repetitive immunization multiple sites strategy with keyhole limpet hemocyanin-conjugated peptides derived from the C5aR, either alone (designated as C5aR-P1 [aa 1–21] and C5aR-P2 [aa 19–31]) or in combination (designated as C5aR-P1+C5aR-P2). Mice were fed a high-fat diet for 10 weeks. Lesions were evaluated histologically; local and systemic immune responses were analyzed by immunohistochemistry of aorta samples and cytokine measurements in plasma samples and splenocyte supernatants. Immunization of
Ldlr

tm1Her

Apob

tm2Sgy

J mice with these peptides elicited high concentrations of antibodies against each peptide. Immunization with the single peptide inhibited plaque development. Combined inoculation with C5aR-P1+C5aR-P2 had an additive effect on reducing the lesion in the aorta sinus and descending aortas when compared with controls. This effect correlated with cellular infiltration and cytokine/chemokine secretion in the serum or in stimulated spleen cells as well as specific cellular immune responses when compared with controls.

Conclusion—

Immunization of mice with C5aR-P1 and C5aR-P2, either alone or in combination, was effective in reducing early atherosclerotic lesion development. The combined peptide is more potential than either epitope alone to reduce atherosclerotic lesion formation through the induction of a specific Treg cell response as well as blockage of monocyte differentiation into macrophages.
Two Types of Procoagulant Platelets Are Formed Upon Physiological Activation and Are Controlled by Integrin α
IIb
β
3
Authors: Topalov Nikolay N. , Yakimenko Alena O. , Canault Matthias , Artemenko Elena O. , Zakharova Natalia V. , Abaeva Anastasia A. , Loosveld Marie , Ataullakhanov Fazoil I. , Nurden Alan T. , Alessi Marie-Christine , Panteleev Mikhail A.
Published: July 2012

Objective—

Phosphatidylserine (PS) externalization by platelets upon activation is a key event in hemostasis and thrombosis. It is currently believed that strong stimulation of platelets forms 2 subpopulations, only 1 of which expresses PS.

Methods and Results—

Here, we demonstrate that physiological stimulation leads to the formation of not 1 but 2 types of PS-expressing activated platelets, with dramatically different properties. One subpopulation sustained increased calcium level after activation, whereas another returned to the basal low-calcium state. High-calcium PS-positive platelets had smaller size, high surface density of fibrin(ogen), no active integrin α
IIb
β
3
, depolarized mitochondrial membranes, gradually lost cytoplasmic membrane integrity, and were poorly aggregated. In contrast, the low-calcium PS-positive platelets had normal size, retained mitochondrial membrane potential and cytoplasmic membrane integrity, and combined retention of fibrin(ogen) with active α
IIb
β
3
and high proaggregatory function. Formation of low-calcium PS-positive platelets was promoted by platelet concentration increase or shaking and was decreased by integrin α
IIb
β
3
antagonists, platelet dilution, or in platelets from kindlin-3–deficient and Glanzmann thrombasthenia patients.

Conclusion—

Identification of a novel PS-expressing platelet subpopulation with low calcium regulated by integrin α
IIb
β
3
can be important for understanding the mechanisms of PS exposure and thrombus formation.
Platelets Derived From the Bone Marrow of Diabetic Animals Show Dysregulated Endoplasmic Reticulum Stress Proteins That Contribute to Increased Thrombosis
Authors: Hernández Vera Rodrigo , Vilahur Gemma , Ferrer-Lorente Raquel , Peña Esther , Badimon Lina
Published: July 2012

Objective—

Patients with diabetes mellitus have an increased risk of suffering atherothrombotic syndromes and are prone to clustering cardiovascular risk factors. However, despite their dysregulated glucose metabolism, intensive glycemic control has proven insufficient to reduce thrombotic complications. Therefore, we aimed to elucidate the determinants of thrombosis in a model of type 2 diabetes mellitus with cardiovascular risk factors clustering.

Methods and Results—

Intravital microscopy was used to analyze thrombosis in vivo in Zucker diabetic fatty rats (ZD) and lean normoglycemic controls. Bone marrow (BM) transplants were performed to test the contribution of each compartment (blood or vessel wall) to thrombogenicity. ZD showed significantly increased thrombosis compared with lean normoglycemic controls. BM transplants demonstrated the key contribution of the hematopoietic compartment to increased thrombogenicity. Indeed, lean normoglycemic controls transplanted with ZD-BM showed increased thrombosis with normal glucose levels, whereas ZD transplanted with lean normoglycemic controls–BM showed reduced thrombosis despite presenting hyperglycemia. Significant alterations in megakaryopoiesis and platelet–endoplasmic reticulum stress proteins, protein disulfide isomerase and 78-kDa glucose-regulated protein, were detected in ZD, and increased tissue factor procoagulant activity was detected in plasma and whole blood of ZD.

Conclusion—

Our results indicate that diabetes mellitus with cardiovascular risk factor clustering favors BM production of hyperreactive platelets with altered protein disulfide isomerase and 78-kDa glucose-regulated protein expression that can contribute to increase thrombotic risk independently of blood glucose levels.